Enhancer of zeste homolog 2 inhibitors

ABSTRACT

This invention relates to novel compounds according to Formula (I) which are inhibitors of Enhancer of Zeste Homolog 2 (EZH2), to pharmaceutical compositions containing them, to processes for their preparation, and to their use in therapy for the treatment of cancers.

FIELD OF THE INVENTION

This invention relates to compounds which inhibit Enhancer of ZesteHomolog 2 (EZH2) and thus are useful for inhibiting the proliferation ofand/or inducing apoptosis in cancer cells.

BACKGROUND OF THE INVENTION

Epigenetic modifications play an important role in the regulation ofmany cellular processes including cell proliferation, differentiation,and cell survival. Global epigenetic modifications are common in cancer,and include global changes in DNA and/or histone methylation,dysregulation of non-coding RNAs and nucleosome remodeling leading toaberrant activation or inactivation of oncogenes, tumor suppressors andsignaling pathways. However, unlike genetic mutations which arise incancer, these epigenetic changes can be reversed through selectiveinhibition of the enzymes involved. Several methylases involved inhistone or DNA methylation are known to be dysregulated in cancer. Thus,selective inhibitors of particular methylases will be useful in thetreatment of proliferative diseases such as cancer.

EZH2 (human EZH2 gene: Cardoso, C, et al; European J of Human Genetics,Vol. 8, No. 3 Pages 174-180, 2000) is the catalytic subunit of thePolycomb Repressor Complex 2 (PRC2) which functions to silence targetgenes by tri-methylating lysine 27 of histone H3 (H3K27me3). Histone H3is one of the five main histone proteins involved in the structure ofchromatin in eukaryotic cells. Featuring a main globular domain and along N-terminal tail, Histones are involved with the structure of thenucleosomes, a ‘beads on a string’ structure. Histone proteins arehighly post-translationally modified however Histone H3 is the mostextensively modified of the five histones. The term “Histone H3” aloneis purposely ambiguous in that it does not distinguish between sequencevariants or modification state. Histone H3 is an important protein inthe emerging field of epigenetics, where its sequence variants andvariable modification states are thought to play a roR^(c) in thedynamic and long term regulation of genes.

Increased EZH2 expression has been observed in numerous solid tumorsincluding those of the prostate, breast, skin, bladder, liver, pancreas,head and neck and correlates with cancer aggressiveness, metastasis andpoor outcome (Varambally et al., 2002; Kleer et al., 2003; Breuer etal., 2004; Bachmann et al., 2005; Weikert et al., 2005; Sudo et al.,2005; Bachmann et al., 2006). For instance, there is a greater risk ofrecurrence after prostatectomy in tumors expressing high levels of EZH2,increased metastasis, shorter disease-free survival and increased deathin breast cancer patients with high EZH2 levels (Varambally et al.,2002; Kleer et al., 2003). More recently, inactivating mutations in UTX(ubiquitously transcribed tetratricopeptide repeats X), a H3K27demethylase which functions in opposition to EZH2, have been identifiedin multiple solid and hematological tumor types (including renal,glioblastoma, esophageal, breast, colon, non-small cell lung, small celllung, bladder, multiple myeloma, and chronic myeloid leukemia tumors),and low UTX levels correlate with poor survival in breast cancersuggesting that loss of UTX function leads to increased H3K27me3 andrepression of target genes (Wang et al., 2010). Together, these datasuggest that increased H3K27me3 levels contribute to canceraggressiveness in many tumor types and that inhibition of EZH2 activitymay provide therapeutic benefit.

Numerous studies have reported that direct knockdown of EZH2 via siRNAor shRNA or indirect loss of EZH2 via treatment with the SAH hydrolaseinhibitor 3-deazaneplanocin A (DZNep) decreases cancer cell lineproliferation and invasion in vitro and tumor growth in vivo (Gonzalezet al., 2008, GBM 2009). WhiR^(c) the precise mechanism by whichaberrant EZH2 activity leads to cancer progression is not known, manyEZH2 target genes are tumor suppressors suggesting that loss of tumorsuppressor function is a key mechanism. In addition, EZH2 overexpressionin immortalized or primary epithelial cells promotes anchorageindependent growth and invasion and requires EZH2 catalytic activity(Kleer et al., 2003; Cao et al., 2008).

Thus, there is strong evidence to suggest that inhibition of EZH2activity decreases cellular proliferation and invasion. Accordingly,compounds that inhibit EZH2 activity would be useful for the treatmentof cancer.

SUMMARY OF THE INVENTION

The present invention relates to compounds according to Formula (I):

wherein:

A is O or NH;

X is O, N, S, CR⁶, or NR⁷;

Y is O, N, S, CR⁶, or NR⁷;

Z is CR⁵ or NR⁸; wherein when X is O, S, or NR⁷, Y is N or CR⁶ and Z isCR⁵; when Y is O, S, or NR⁷, X is N or CR⁶ and Z is CR⁵; and when Z isNR⁸, Y is N or CR⁶ and X is N or CR⁶;

R¹, R², and R³ are each independently selected from the group consistingof hydrogen, (C₁-C₄)alkoxy, (C₁-C₈)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl-,halo(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, hydroxy(C₁-C₄)alkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, R^(a)O(O)CNH(C₁-C₄)alkyl-,(C₆-C₁₀)bicycloalkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-,aryl, aryl(C₁-C₄)alkyl, heteroaryl, heteroaryl(C₁-C₄)alkyl, halogen,cyano, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(b),—S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b), nitro, —NR^(a)R^(b),—NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a),—NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b),—NR^(a)NR^(a)C(O)R^(b), —NR^(a)NR^(a)C(O)NR^(a)R^(b),—NR^(a)NR^(a)C(O)OR^(a), —OR^(a), —OC(O)R^(a), and —OC(O)NR^(a)R^(b),wherein each (C₃-C₈)cycloalkyl, (C6-C₁₀)bicycloalkyl, heterocycloalkyl,aryl, or heteroaryl is optionally substituted 1, 2, or 3 times,independently, by hydroxyl, halogen, nitro, (C₁-C₄)alkyl, cyano,(C₁-C₄)alkoxy, —NR^(a)R^(b) or —CO₂R^(a);

R⁴ is selected from the group consisting of hydrogen, (C₁-C₃)alkoxy,(C₁-C₃)alkyl, hydroxyl, halogen, cyano, (C₃-C₆)cycloalkyl,heterocycloalkyl, —NR^(a)R^(b), halo(C₁-C₃)alkyl, andhydroxy(C₁-C₃)alkyl;

R⁵ is selected from the group consisting of (C₄-C₈)alkyl,(C₂-C₈)alkenyl, (C₃-C₈)alkoxy, (C₄-C₈)cycloalkyl,(C₃-C₈)cycloalkyl(C₁-C₂)alkyl-, (C₃-C₈)cycloalkyloxy-, heterocycloalkyl,heterocycloalkyl(C₁-C₂)alkyl-, heterocycloalkyloxy-, aryl, heteroaryl,and —NR^(a)R^(b), wherein said (C₄-C₈)alkyl, (C₂-C₈)alkenyl,(C₃-C₈)alkoxy, (C₄-C₈)cycloalkyl, (C3-C8)cycloalkyl(C₁-C₂)alkyl-,(C₃-C₈)cycloalkyloxy-, heterocycloalkyl, heterocycloalkyl(C₁-C₂)alkyl-,heterocycloalkyloxy-, aryl, or heteroaryl is optionally substituted 1,2, or 3 times, independently, by halogen, —OR^(a), —NR^(a)R^(b),—NHCO₂R^(a), nitro, (C₁-C₃)alkyl, R^(a)R^(b)N(C₁-C₃)alkyl-,R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano, —CO₂R^(a),—C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), heterocycloalkyl, aryl, orheteroaryl, wherein said (C3-C8)cycloalkyl, heterocycloalkyl, aryl, orheteroaryl is optionally substituted 1 or 2 times, independently, byhalogen, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, —CO(C₁-C₄)alkyl,—CO₂(C₁-C₄)alkyl, —NR^(a)R^(b), —NHCO₂R^(a), hydroxyl, oxo,(C₁-C₄)alkoxy, or (C₁-C₄)alkoxy(C₁-C₄)alkyl-; or any 2 optionalsubstituents on said (C₂-C₈)alkenyl taken together with the carbonatom(s) to which they are attached represent a 5-8 membered ring,optionally containing a heteroatom selected from oxygen, nitrogen, andsulfur, wherein said ring is optionally substituted 1 or 2 times,independently, by (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, —CO(C₁-C₄)alkyl,—CO₂(C₁-C₄)alkyl, —NR^(a)R^(b), —NHCO₂R^(a), hydroxyl, oxo,(C₁-C₄)alkoxy, or (C₁-C₄)alkoxy(C₁-C₄)alkyl-;

R⁶ is selected from the group consisting of hydrogen, halogen,(C₁-C₈)alkyl, (C₁-C4)alkoxy, —B(OH)₂, (C₃-C₈)cycloalkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C6-C₁₀)bicycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl, heteroaryl,heteroaryl(C₁-C₄)alkyl, cyano, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b),—C(O)NR^(a)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), R^(a)R^(b)N(C₁-C₄)alkyl-, —NR^(a)C(O)R^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b),—NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b), —NR^(a)NR^(a)C(O)R^(b),—NR^(a)NR^(a)C(O)NR^(a)R^(b), —NR^(a)NR^(a)C(O)OR^(a), —OR^(a),—OC(O)R^(a), and —OC(O)NR^(a)R^(b), wherein each cycloalkyl,bicycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is optionallysubstituted 1, 2, or 3 times, independently, by R^(c)-(C₁-C₆)alkyl-O—,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heterocycloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl;

R⁷ is selected from the group consisting of hydrogen, (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl, (C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C₆-C₁₀)bicycloalkyl,heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl,heteroaryl, heteroaryl(C₁-C₄)alkyl, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(a)R^(b), —SO₂R^(a), —SO₂NR^(a)R^(b),and R^(a)R^(b)N(C₁-C₄)alkyl-, wherein each cycloalkyl, bicycloalkyl,heterocycloalkyl, aryl, or heteroaryl group is optionally substituted 1,2, or 3 times, independently, by R^(c)-(C₁-C₆)alkyl-O—,R^(c)-(C₁-C₆)alkyl-S—, R^(c)-(C₁-C₆)alkyl-,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heterocycloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl;

R⁸ is selected from the group consisting of (C₄-C₈)alkyl,(C₄-C₈)cycloalkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₂)alkyl-,aryl, and heteroaryl, wherein said (C4-C8)alkyl, (C₄-C₈)cycloalkyl,heterocycloalkyl, heterocycloalkyl(C₁-C₂)alkyl-, aryl, or heteroaryl isoptionally substituted 1, 2, or 3 times, independently, by halogen,—OR^(a), —NR^(a)R^(b), —NHCO₂R^(a), nitro, (C₁-C₃)alkyl,R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano,—CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), aryl, or heteroaryl;

each R^(c) is independently —S(O)R^(a), —SO₂R^(a), —NR^(a)R^(b)—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), or —CO₂R^(a); and

R^(a) and R^(b) are each independently hydrogen, (C₁-C₄)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl-, (C₃-C₁₀)cycloalkyl, heterocycloalkyl, aryl,aryl(C₁-C₄)alkyl-, heteroaryl(C₁-C₄)alkyl-, or heteroaryl, wherein anysaid cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group isoptionally substituted 1, 2, or 3 times, independently, by halogen,hydroxyl, (C₁-C₄)alkoxy, amino, —NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, —CO₂H, —CO₂(C₁-C₄)alkyl, —CONH₂,—CONH(C₁-C₄)alkyl, —CON((C₁-C₄)alkyl)₂, —SO₂(C₁-C₄)alkyl, —SO₂NH₂,—SO₂NH(C₁-C₄)alkyl, or —SO₂N((C₁-C₄)alkyl)₂;

or R^(a) and R^(b) taken together with the nitrogen to which they areattached represent a 5-8 membered saturated or unsaturated ring,optionally containing an additional heteroatom selected from oxygen,nitrogen, and sulfur, wherein said ring is optionally substituted 1, 2,or 3 times, independently, by (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, amino,—NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂, hydroxyl, oxo, (C₁-C₄)alkoxy, or(C₁-C₄)alkoxy(C₁-C₄)alkyl-, wherein said ring is optionally fused to a(C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring;

or R^(a) and R^(b) taken together with the nitrogen to which they areattached represent a 6- to 10-membered bridged bicyclic ring systemoptionally fused to a (C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, orheteroaryl ring;

or pharmaceutically acceptable salts thereof.

Another aspect of this invention relates to a method of inducingapoptosis in cancer cells of solid tumors; treating solid tumor cancers.

Another aspect of the invention relates to pharmaceutical preparationscomprising compounds of Formula (I) and pharmaceutically acceptableexcipients.

In another aspect, there is provided the use of a compound of Formula(I) and/or a pharmaceutically acceptable salt or solvate thereof, in thepreparation of a medicament for use in the treatment of a disordermediated by EZH2, such as by inducing apoptosis in cancer cells.

In another aspect, this invention provides for the use of a compound ofFormula (I) or a pharmaceutically acceptable salt thereof for thetreatment of diseases mediated by EZH2. The invention further providesfor the use of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof as an active therapeutic substance in thetreatment of a disease mediated by EZH2.

In another aspect, the invention provides a compound of Formula (I) or apharmaceutically acceptable salt thereof for use in therapy.

In another aspect there is provided methods of co-administering thepresently invented compounds of Formula (I) with other activeingredients.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to compounds of the Formula (I) as defined above.

In one embodiment, this invention relates to compounds of Formula (I),wherein:

A is O or NH;

X is O, N, S, CR⁶, or NR⁷;

Y is O, N, S, CR⁶, or NR⁷;

Z is CR⁵ or NR⁸; wherein when X is O, S, or NR⁷, Y is N or CR⁶ and Z isCR⁵; when Y is O, S, or NR⁷, X is N or CR⁶ and Z is CR⁵; and when Z isNR⁸, Y is N or CR⁶ and X is N or CR⁶;

R¹, R², and R³ are each independently selected from the group consistingof hydrogen, (C₁-C₄)alkoxy, (C₁-C₈)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl-,halo(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, hydroxy(C₁-C₄)alkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, R^(a)O(O)CNH(C₁-C₄)alkyl-,(C₆-C₁₀)bicycloalkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-,aryl, aryl(C₁-C₄)alkyl, heteroaryl, heteroaryl(C₁-C₄)alkyl, halogen,cyano, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(a)R^(b),—S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b), nitro, —NR^(a)R^(b),—NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a),—NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b),—NR^(a)NR^(a)C(O)R^(b), —NR^(a)NR^(a)C(O)NR^(a)R^(b),—NR^(a)NR^(a)C(O)OR^(a), —OR^(a), —OC(O)R^(a), and —OC(O)NR^(a)R^(b),wherein each (C₃-C₈)cycloalkyl, (C6-C₁₀)bicycloalkyl, heterocycloalkyl,aryl, or heteroaryl is optionally substituted 1, 2, or 3 times,independently, by hydroxyl, halogen, nitro, (C₁-C₄)alkyl, cyano,(C₁-C₄)alkoxy, —NR^(a)R^(b) or —CO₂R^(a);

R⁴ is selected from the group consisting of hydrogen, (C₁-C₃)alkoxy,(C₁-C₃)alkyl, hydroxyl, halogen, cyano, (C₃-C₆)cycloalkyl,heterocycloalkyl, —NR^(a)R^(b), halo(C₁-C₃)alkyl, andhydroxy(C₁-C₃)alkyl;

R⁵ is selected from the group consisting of (C₄-C₈)alkyl, (C₃-C₈)alkoxy,(C₄-C₈)cycloalkyl, (C₃-C₈)cycloalkyloxy-, heterocycloalkyl,heterocycloalkyloxy-, aryl, heteroaryl, and —NR^(a)R^(b), wherein said(C₄-C₈)alkyl, (C₃-C₈)alkoxy, (C₄-C₈)cycloalkyl, (C3-C8)cycloalkyloxy-,heterocycloalkyl, heterocycloalkyloxy-, aryl, or heteroaryl isoptionally substituted 1, 2, or 3 times, independently, by halogen,—OR^(a), —NR^(a)R^(b), —NHCO₂R^(a), nitro, (C₁-C₃)alkyl,R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano,—CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), aryl, or heteroaryl;

R⁶ is selected from the group consisting of hydrogen, halogen,(C₁-C₈)alkyl, (C₁-C4)alkoxy, —B(OH)₂, (C₃-C₈)cycloalkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C6-C₁₀)bicycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl, heteroaryl,heteroaryl(C₁-C₄)alkyl, cyano, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b),—C(O)NR^(a)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), R^(a)R^(b)N(C₁-C₄)alkyl-, —NR^(a)C(O)R^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b),—NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b), —NR^(a)NR^(a)C(O)R^(b),—NR^(a)NR^(a)C(O)NR^(a)R^(b), —NR^(a)NR^(a)C(O)OR^(a), —OR^(a),—OC(O)R^(a), and —OC(O)NR^(a)R^(b), wherein each cycloalkyl,bicycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is optionallysubstituted 1, 2, or 3 times, independently, by R° -(C₁-C₆)alkyl-O—,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b), nitro,—NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heterocycloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl;

R⁷ is selected from the group consisting of hydrogen, (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl, (C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C₆-C₁₀)bicycloalkyl,heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl,heteroaryl, heteroaryl(C₁-C₄)alkyl, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(a)R^(b), —SO₂R^(a), —SO₂NR^(a)R^(b),and R^(a)R^(b)N(C₁-C₄)alkyl-, wherein each cycloalkyl, bicycloalkyl,heterocycloalkyl, aryl, or heteroaryl group is optionally substituted 1,2, or 3 times, independently, by R^(c)-(C₁-C₆)alkyl-O—,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b), nitro,—NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heterocycloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl;

R⁸ is selected from the group consisting of (C₄-C₈)alkyl,(C₄-C₈)cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, wherein said(C₄-C₈)alkyl, (C₄-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroarylis optionally substituted 1, 2, or 3 times, independently, by halogen,—OR^(a), —NR^(a)R^(b), —NHCO₂R^(a), nitro, (C₁-C₃)alkyl,R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano,—CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), aryl, or heteroaryl;

each R^(c) is independently —S(O)R^(a), —SO₂R^(a), —NR^(a)R^(b)—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), or —CO₂R^(a); and

R^(a) and R^(b) are each independently hydrogen, (C₁-C₄)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl-, (C₃-C₁₀)cycloalkyl, heterocycloalkyl, aryl,aryl(C₁-C₄)alkyl-, heteroaryl(C₁-C₄)alkyl-, or heteroaryl, wherein anysaid cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group isoptionally substituted 1, 2, or 3 times, independently, by halogen,hydroxyl, (C₁-C₄)alkoxy, amino, —NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, —CO₂H, —CO₂(C₁-C₄)alkyl, —CONH₂,—CONH(C₁-C₄)alkyl, —CON((C₁-C₄)alkyl)₂, —SO₂(C₁-C₄)alkyl, —SO₂NH₂,—SO₂NH(C₁-C₄)alkyl, or —SO₂N((C₁-C₄)alkyl)₂;

or R^(a) and R^(b) taken together with the nitrogen to which they areattached represent a 5-8 membered saturated or unsaturated ring,optionally containing an additional heteroatom selected from oxygen,nitrogen, and sulfur, wherein said ring is optionally substituted 1, 2,or 3 times, independently, by (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, amino,—NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂, hydroxyl, oxo, (C₁-C₄)alkoxy, or(C₁-C₄)alkoxy(C₁-C₄)alkyl-, wherein said ring is optionally fused to a(C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring;

or R^(a) and R^(b) taken together with the nitrogen to which they areattached represent a 6- to 10-membered bridged bicyclic ring systemoptionally fused to a (C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, orheteroaryl ring; or pharmaceutically acceptable salts thereof.

In one embodiment, this invention relates to compounds of Formula (I),wherein A is NH. In another embodiment, this invention relates tocompounds of Formula (I), wherein A is O.

In one embodiment, this invention relates to compounds of Formula (I),wherein X is O, S, or NR⁷; Y is N or CR⁶; and Z is CR⁵. In anotherembodiment, this invention relates to compounds of Formula (I), whereinX is O or S; Y is N or CR⁶; and Z is CR⁵. In another embodiment, thisinvention relates to compounds of Formula (I), wherein X is O or S; Y isCR⁶; and Z is CR⁵. In another embodiment, this invention relates tocompounds of Formula (I), wherein X is O or S; Y is N; and Z is CR⁵. Inanother embodiment, this invention relates to compounds of Formula (I),wherein X is S; Y is CR⁶; and Z is CR⁵. In another embodiment, thisinvention relates to compounds of Formula (I), wherein Y is O, S, orNR⁷; X is N or CR⁶; and Z is CR⁵. In another embodiment, this inventionrelates to compounds of Formula (I), wherein Y is O or S; X is N or CR⁶;and Z is CR⁵. In another embodiment, this invention relates to compoundsof Formula (I), wherein Y is O or S; X is CR⁶; and Z is CR⁵. In anotherembodiment, this invention relates to compounds of Formula (I), whereinY is O or S; X is N; and Z is CR⁵. In another embodiment, this inventionrelates to compounds of Formula (I), wherein Y is S; X is CR⁶; and Z isCR⁵. In another embodiment, this invention relates to compounds ofFormula (I), wherein Z is NR⁸; Y is N or CR⁶; and X is N or CR⁶.

In another embodiment, this invention relates to compounds of Formula(I), wherein R¹, R², and R³ are each independently selected from thegroup consisting of hydrogen, (C₁-C₄)alkoxy, (C₁-C₄)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl-, halo(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl,hydroxy(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl(C₁-C₄)alkyl-,(C₁-C₄)alkylO(O)CNH(C₁-C₄)alkyl-, amino, —NH(C₁-C₄)alkyl,—N((C₁-C₄)alkyl)₂, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-,aryl, aryl(C₁-C₄)alkyl-, heteroaryl, and heteroaryl(C₁-C₄)alkyl-,wherein each (C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroaryl isoptionally substituted 1 or 2 times, independently, by hydroxyl,halogen, nitro, (C₁-C₄)alkyl, cyano, (C₁-C₄)alkoxy, —NH(C₁-C₄)alkyl,—N((C₁-C₄)alkyl)₂, or —CO₂(C₁-C₄)alkyl. In another embodiment, thisinvention relates to compounds of Formula (I), wherein R¹, R², and R³are each independently selected from the group consisting of hydrogen,(C₁-C₄)alkoxy, (C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl-,halo(C₁-C₄)alkyl, hydroxy(C₁-C₄)alkyl, amino, —NH(C₁-C₄)alkyl, and—N((C₁-C₄)alkyl)₂.

In another embodiment, this invention relates to compounds of Formula(I), wherein R^(c) and R² are each independently (C₁-C₄)alkyl.

In a specific embodiment, this invention relates to compounds of Formula(I), wherein R¹ is methyl.

In another specific embodiment, this invention relates to compounds ofFormula (I), wherein R² is methyl.

In another specific embodiment, this invention relates to compounds ofFormula (I), wherein R¹ and R² are each methyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R³ is hydrogen, amino, —NH(C₁-C₄)alkyl, or—N((C₁-C₄)alkyl)₂. In a specific embodiment, this invention relates tocompounds of Formula (I), wherein R³ is amino. In another specificembodiment, this invention relates to compounds of Formula (I), whereinR³ is hydrogen.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁴ is selected from the group consisting of hydrogen,(C₁-C₃)alkyl, hydroxyl, halogen, halo(C₁-C₃)alkyl, andhydroxy(C₁-C₃)alkyl. In another embodiment, this invention relates tocompounds of Formula (I), wherein R⁴ is (C₁-C₃)alkyl or halogen. In aspecific embodiment, this invention relates to compounds of Formula (I),wherein R⁴ is methyl or chlorine. In another specific embodiment, thisinvention relates to compounds of Formula (I), wherein R⁴ is methyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is selected from the group consisting of (C₃-C₆)alkoxy,(C₃-C₆)cycloalkyloxy-, heterocycloalkyloxy-, heterocycloalkyl,—NH((C₃-C₆)cycloalkyl), —N((C₁-C₃)alkyl)((C₃-C₆)cycloalkyl),—NH(heterocycloalkyl), and —N((C₁-C₃)alkyl)(heterocycloalkyl), whereinany said (C₃-C₆)alkoxy, (C₃-C₆)cycloalkyloxy-, heterocycloalkyloxy-,heterocycloalkyl, or (C₃-C₆)cycloalkyl is optionally substituted 1 or 2times, independently, by halogen, hydroxyl, (C₁-C₃)alkoxy, amino,—NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂, (C₁-C₃)alkyl,(C₁-C₃)alkoxy(C₁-C₃)alkyl-, amino(C₁-C₃)alkyl-,((C₁-C₃)alkyl)NH(C₁-C₃)alkyl-, ((C₁-C₃)alkyl)₂N(C₁-C₃)alkyl-,(C₃-C₈)cycloalkyl, cyano, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),phenyl, or heteroaryl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is selected from the group consisting of (C₃-C₆)alkoxy,(C₃-C₈)cycloalkyloxy-, and heterocycloalkyloxy-, each of which isoptionally substituted by hydroxyl, (C₁-C₃)alkoxy, amino,—NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂, (C₁-C₃)alkyl, —CO₂R^(a),—C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), phenyl, or heteroaryl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is (C₃-C₆)cycloalkyloxy- which is optionally substituted1, 2, or 3 times, independently, by halogen, —OR⁷, —NR^(a)R^(b), nitro,(C₁-C₃)alkyl, R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-,(C₃-C₈)cycloalkyl, cyano, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),aryl, or heteroaryl. In another embodiment, this invention relates tocompounds of Formula (I), wherein R⁵ is (C₃-C₆)cycloalkyloxy- which isoptionally substituted 1 or 2 times, independently, by halogen,hydroxyl, (C₁-C₃)alkoxy, amino, —NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂,(C₁-C₃)alkyl, (C₁-C₃)alkoxy(C₁-C₃)alkyl-, amino(C₁-C₃)alkyl-,((C₁-C₃)alkyl)NH(C₁-C₃)alkyl-, ((C₁-C₃)alkyl)₂N(C₁-C₃)alkyl-,(C₃-C₈)cycloalkyl, cyano, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),phenyl, or heteroaryl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is heterocycloalkyloxy- which is optionally substituted1, 2, or 3 times, independently, by halogen, —OR^(a), —NR^(a)R^(b),nitro, (C₁-C₃)alkyl, R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-,(C₃-C₈)cycloalkyl, cyano, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),aryl, or heteroaryl. In another embodiment, this invention relates tocompounds of Formula (I), wherein R⁵ is heterocycloalkyloxy- which isoptionally substituted 1 or 2 times, independently, by halogen,hydroxyl, (C₁-C₃)alkoxy, amino, —NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂,(C₁-C₃)alkyl, (C₁-C₃)alkoxy(C₁-C₃)alkyl-, amino(C₁-C₃)alkyl-,((C₁-C₃)alkyl)NH(C₁-C₃)alkyl-, ((C₁-C₃)alkyl)₂N(C₁-C₃)alkyl-,(C₃-C₈)cycloalkyl, cyano, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),_(p)hen_(y)l, or heteroaryl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is selected from the group consisting of cyclopentyloxy,cyclohexyloxy, pyrrolidinyloxy, piperidinyloxy, andtetrahydropyranyloxy, each of which is optionally substituted byhydroxyl, (C₁-C₃)alkoxy, amino, —NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂,(C₁-C₃)alkyl, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),_(p)hen_(y)l, furanyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl,triazolyl, tetrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl,oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrazinyl, orpyrimidinyl, wherein R^(a) is (C₁-C₄)alkyl or phenyl(C₁-C₂)alkyl andR^(b) is hydrogen or (C₁-C₄)alkyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is —NR^(a)R^(b). In another embodiment, this inventionrelates to compounds of Formula (I), wherein R⁵ is —NR^(a)R^(b); R^(a)is azetidinyl, oxetanyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, or tetrahydropyranyl, each of which isoptionally substituted 1 or 2 times, independently, by (C₁-C₄)alkyl; andR^(b) is hydrogen or (C₁-C₄)alkyl. In another embodiment, this inventionrelates to compounds of Formula (I), wherein R⁵ is —NR^(a)R^(b); R^(a)is azetidinyl, oxetanyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, or tetrahydropyranyl; and R^(b) is methylor ethyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁵ is —NR^(a)R^(b); R^(a) is cyclopentyl or cyclohexyl,each of which is optionally substituted by amino, —NH(C₁-C₄)alkyl, or—N((C₁-C₄)alkyl)₂; and R^(b) is hydrogen or (C₁-C₄)alkyl. In anotherembodiment, this invention relates to compounds of Formula (I), whereinR⁵ is —NR^(a)R^(b); R^(a) is cyclopentyl or cyclohexyl, each of which isoptionally substituted by —N((C₁-C₂)alkyl)₂; and R^(b) is methyl orethyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁶ is selected from the group consisting of hydrogen,—SO₂(C₁-C₄)alkyl, halogen, (C₁-C₆)alkyl, (C₁-C₄)alkoxy, phenyl,heteroaryl, and cyano, wherein said phenyl or heteroaryl group isoptionally substituted 1 or 2 times, independently, by (C₁-C₄)alkoxy,—NR^(a)R^(b), leR^(b)N(C₁-C₄)alkyl-, (C₁-C₄)alkylheterocycloalkyl-,halogen, (C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, or heterocycloalkyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁶ is selected from the group consisting of hydrogen,cyano, halogen, (C₁-C₄)alkoxy, furanyl, thienyl, pyrrolyl, imidazolyl,pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl,oxadiazolyl, thiadiazolyl, isothiazolyl, phenyl, pyridinyl, pyridazinyl,pyrazinyl, pyrimidinyl, and triazinyl, wherein said furanyl, thienyl,pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl,oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, phenyl,pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, or triazinyl isoptionally substituted by (C₁-C₄)alkoxy, —NR^(a)R^(b),R^(a)R^(b)N(C₁-C₄)alkyl-, (C₁-C₄)alkylheterocycloalkyl-, halogen,(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, or heterocycloalkyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁶ is phenyl which is optionally substituted by—NR^(a)R^(b) or R^(a)R^(b)N(C₁-C₄)alkyl-.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁶ is pyridinyl which is optionally substituted by—NR^(a)R^(b) or R^(a)R^(b)N(C₁-C₄)alkyl-.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁶ is hydrogen, halogen, (C₁-C₄)alkyl, or (C₁-C₄)alkoxy. Inanother embodiment, this invention relates to compounds of Formula (I),wherein R⁶ is hydrogen or halogen. In a specific embodiment, thisinvention relates to compounds of Formula (I), wherein R⁶ is hydrogen,fluorine, chlorine, or bromine. In a specific embodiment, this inventionrelates to compounds of Formula (I), wherein R⁶ is hydrogen or chlorine.In a more specific embodiment, this invention relates to compounds ofFormula (I), wherein R⁶ is chlorine. In another specific embodiment,this invention relates to compounds of Formula (I), wherein R⁶ ishydrogen.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁷ is selected from the group consisting of hydrogen,(C₁-C₆)alkyl, (C₃-C₆)cycloalkyl, phenyl, and heteroaryl, wherein saidphenyl or heteroaryl group is optionally substituted 1 or 2 times,independently, by (C₁-C₄)alkoxy, —NR^(a)R^(b), R^(a)R^(b)N(C₁-C₄)alkyl-,(C₁-C₄)alkylheterocycloalkyl-, halogen, (C₁-C₄)alkyl, (C₃-C₈)cycloalkyl,or heterocycloalkyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁷ is hydrogen or (C₁-C₄)alkyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁷ is selected from the group consisting of furanyl,thienyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl,thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl,isothiazolyl, phenyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl,and triazinyl, wherein said furanyl, thienyl, pyrrolyl, imidazolyl,pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl,oxadiazolyl, thiadiazolyl, isothiazolyl, phenyl, pyridinyl, pyridazinyl,pyrazinyl, pyrimidinyl, or triazinyl is optionally substituted by(C₁-C₄)alkoxy, —NR^(a)R^(b),

R^(a)R^(b)N(C₁-C₄)alkyl-, (C₁-C₄)alkylheterocycloalkyl-, halogen,(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, or heterocycloalkyl.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁷ is phenyl which is optionally substituted by—NR^(a)R^(b) or R^(a)R^(b)N(C₁-C₄)alkyl-.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁷ is pyridinyl which is optionally substituted by—NR^(a)R^(b) or R^(a)RN(C₁-C₄)alkyl-.

In another embodiment, this invention relates to compounds of Formula(I), wherein R⁸ is selected from the group consisting of (C₄-C₆)alkyl,(C₄-C₆)cycloalkyl, heterocycloalkyl, and phenyl, wherein said(C₄-C₆)alkyl, (C₄-C₆)cycloalkyl, heterocycloalkyl, or phenyl isoptionally substituted 1 or 2 times, independently, by —OR^(a),—NR^(a)R^(b), —NHCO₂R^(a), (C₁-C₃)alkyl, R^(a)R^(b)N(C₁-C₃)alkyl-,R^(a)O(C₁-C₃)alkyl-, —CO₂R^(a), —C(O)NR^(a)R^(b), or —SO₂NR^(a)R^(b).

In a particular embodiment, this invention relates to compounds ofFormula (I), wherein:

X is O, S, or NR⁷;

Y is N or CR⁶;

Z is CR⁵;

R¹ and R² are each independently (C₁-C₄)alkyl;

R³ is hydrogen;

R⁴ is methyl or chlorine;

R⁵ is selected from the group consisting of (C₃-C₆)alkoxy,(C₃-C₈)cycloalkyloxy-, and heterocycloalkyloxy-, each of which isoptionally substituted by hydroxyl, (C₁-C₃)alkoxy, amino,—NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂, (C₁-C₃)alkyl, —CO₂R^(a),—C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), phenyl, or heteroaryl;

R⁶ is hydrogen, halogen, (C₁-C₄)alkyl, or (C₁-C₄)alkoxy; and

R⁷ is hydrogen or (C₁-C₄)alkyl;

or pharmaceutically acceptable salts thereof.

In another particular embodiment, this invention relates to compounds ofFormula (I), wherein:

X is O, S, or NR⁷;

Y is N or CR⁶;

Z is CR⁵;

R¹ and R² are each independently (C₁-C₄)alkyl;

R³ is hydrogen;

R⁴ is methyl or chlorine;

R⁵ is —NR^(a)R^(b);

R⁶ is hydrogen, halogen, (C₁-C₄)alkyl, or (C₁-C₄)alkoxy; and

R⁷ is hydrogen or (C₁-C₄)alkyl;

or pharmaceutically acceptable salts thereof.

In another particular embodiment, this invention relates to compounds ofFormula (I), wherein:

X is N or CR⁶;

Y is O, S, or NR⁷;

Z is CR⁵;

R¹ and R² are each independently (C₁-C₄)alkyl;

R³ is hydrogen;

R⁴ is methyl or chlorine;

R⁵ is selected from the group consisting of (C₃-C₆)alkoxy,(C₃-C₈)cycloalkyloxy-, and heterocycloalkyloxy-, each of which isoptionally substituted by hydroxyl, (C₁-C₃)alkoxy, amino,—NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂, (C₁-C₃)alkyl, —CO₂R^(a),—C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), phenyl, or heteroaryl;

R⁶ is hydrogen, halogen, (C₁-C₄)alkyl, or (C₁-C₄)alkoxy; and

R⁷ is hydrogen or (C₁-C₄)alkyl;

or pharmaceutically acceptable salts thereof.

In another particular embodiment, this invention relates to compounds ofFormula (I), wherein:

X is N or CR⁶;

Y is O, S, or NR⁷;

Z is CR⁵;

R¹ and R² are each independently (C₁-C₄)alkyl;

R³ is hydrogen;

R⁴ is methyl or chlorine;

R⁵ is —NR^(a)R^(b);

R⁶ is hydrogen, halogen, (C₁-C₄)alkyl, or (C₁-C₄)alkoxy; and

R⁷ is hydrogen or (C₁-C₄)alkyl;

or pharmaceutically acceptable salts thereof.

In another embodiment, this invention also relates to compounds ofFormula (II):

or pharmaceutically acceptable salts thereof, wherein X is O, S, or NR⁷;Y is N or CR⁶; and R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are defined accordingto Formula (I). In another embodiment, this invention relates tocompounds of Formula (II), wherein X is O or S and Y is N or CR⁶. Inanother embodiment, this invention relates to compounds of Formula (II),wherein X is O or S and Y is CR⁶. In another embodiment, this inventionrelates to compounds of Formula (II), wherein X is O or S and Y is N. Inanother embodiment, this invention relates to compounds of Formula (II),wherein X is S and Y is CR⁶.

In another embodiment, this invention also relates to compounds ofFormula (II)(a):

or pharmaceutically acceptable salts thereof, wherein R¹,_(R)2_(, R)3_(,) R4, R5, and R⁶ are defined according to Formula (I).

In another embodiment, this invention also relates to compounds ofFormula (III):

or pharmaceutically acceptable salts thereof, wherein Y is O, S, or NR⁷;X is N or CR⁶; and R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are defined accordingto Formula (I). In another embodiment, this invention relates tocompounds of Formula (III), wherein Y is O or S and X is N or CR⁶. Inanother embodiment, this invention relates to compounds of Formula(III), wherein Y is O or S and X is CR⁶. In another embodiment, thisinvention relates to compounds of Formula (III), wherein Y is O or S andX is N. In another embodiment, this invention relates to compounds ofFormula (III), wherein Y is S and Xis CR⁶.

In another embodiment, this invention also relates to compounds ofFormula (III)(a):

or pharmaceutically acceptable salts thereof, wherein R¹,_(R)2_(, R)3_(,) R4, R5, and R⁶ are defined according to Formula (I).

In another embodiment, this invention also relates to compounds ofFormula (IV):

or pharmaceutically acceptable salts thereof, wherein X is N or CR⁶; Yis N or CR⁶; and R¹, R², R³, R⁴, R⁶, and R⁸ are defined according toFormula (I). In another embodiment, this invention relates to compoundsof Formula (IV), wherein X is N and Y is CR⁶. In another embodiment,this invention relates to compounds of Formula (IV), wherein X is CR⁶and Y is N. In another embodiment, this invention relates to compoundsof Formula (IV), wherein X and Y are each independently CR⁶. In anotherembodiment, this invention relates to compounds of Formula (IV), whereinX and Y are each N.

Specific compounds of this invention include:

N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxamide;

N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide;

N-((5-amino-2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide;and

2-bromo—N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide;

or pharmaceutically acceptable salts thereof.

Typically, but not absolutely, the salts of the present invention arepharmaceutically acceptable salts. Salts of the disclosed compoundscontaining a basic amine or other basic functional group may be preparedby any suitable method known in the art, including treatment of the freebase with an inorganic acid, such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, and the like, or withan organic acid, such as acetic acid, trifluoroacetic acid, maleic acid,succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid,oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid, such asglucuronic acid or galacturonic acid, alpha-hydroxy acid, such as citricacid or tartaric acid, amino acid, such as aspartic acid or glutamicacid, aromatic acid, such as benzoic acid or cinnamic acid, sulfonicacid, such as p-toluenesulfonic acid, methanesulfonic acid,ethanesulfonic acid or the like. Examples of pharmaceutically acceptablesalts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,phosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates succinates, suberates,sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates,hydroxybenzoates, methoxybenzoates, phthalates, phenylacetates,phenylpropionates, phenylbutrates, citrates, lactates,γ-hydroxybutyrates, glycolates, tartrates mandelates, and sulfonates,such as xylenesulfonates, methanesulfonates, propanesulfonates,naphthalene-1-sulfonates and naphthalene-2-sulfonates.

Salts of the disclosed compounds containing a carboxylic acid or otheracidic functional group can be prepared by reacting with a suitablebase. Such a pharmaceutically acceptable salt may be made with a basewhich affords a pharmaceutically acceptable cation, which includesalkali metal salts (especially sodium and potassium), alkaline earthmetal salts (especially calcium and magnesium), aluminum salts andammonium salts, as well as salts made from physiologically acceptableorganic bases such as trimethylamine, triethylamine, morpholine,pyridine, piperidine, picoline, dicyclohexylamine,N,N′-dibenzylethylenediamine, 2-hydroxyethylamine,bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine,dibenzylpiperidine, dehydroabietylamine, N,N′-bisdehydroabietylamine,glucamine, N-methylglucamine, collidine, quinine, quinoline, and basicamino acid such as lysine and arginine.

Other salts, which are not pharmaceutically acceptable, may be useful inthe preparation of compounds of this invention and these should beconsidered to form a further aspect of the invention. These salts, suchas oxalic or trifluoroacetate, while not in themselves pharmaceuticallyacceptable, may be useful in the preparation of salts useful asintermediates in obtaining the compounds of the invention and theirpharmaceutically acceptable salts.

The compound of Formula (I) or a salt thereof may exist instereoisomeric forms (e.g., it contains one or more asymmetric carbonatoms). The individual stereoisomers (enantiomers and diastereomers) andmixtures of these are included within the scope of the presentinvention. Likewise, it is understood that a compound or salt of Formula(I) may exist in tautomeric forms other than that shown in the formulaand these are also included within the scope of the present invention.It is to be understood that the present invention includes allcombinations and subsets of the particular groups defined hereinabove.The scope of the present invention includes mixtures of stereoisomers aswell as purified enantiomers or enantiomerically/diastereomericallyenriched mixtures. It is to be understood that the present inventionincludes all combinations and subsets of the particular groups definedhereinabove.

The subject invention also includes isotopically-labeled compounds,which are identical to those recited in Formula (I) and following, butfor the fact that one or more atoms are replaced by an atom having anatomic mass or mass number different from the atomic mass or mass numberusually found in nature. Examples of isotopes that can be incorporatedinto compounds of the invention and pharmaceutically acceptable saltsthereof include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, sulfur, fluorine, chlorine, and iodine, such as ²H, ³H,¹¹c, ¹³c, ¹⁴c, ¹⁵N, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I.

Compounds of the present invention and pharmaceutically acceptable saltsof said compounds that contain the aforementioned isotopes and/or otherisotopes of other atoms are within the scope of the present invention.Isotopically-labeled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H, ¹⁴C are incorporated,are useful in drug and/or substrate tissue distribution assays.Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes are particularlypreferred for their ease of preparation and detectability. ¹¹C and ¹⁸Fisotopes are particularly useful in PET (positron emission tomography),and ¹²⁵I isotopes are particularly useful in SPECT (single photonemission computerized tomography), all useful in brain imaging. Further,substitution with heavier isotopes such as deuterium, i.e., ²H, canafford certain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements and, hence, may be preferred in some circumstances.Isotopically labeled compounds of Formula (I) and following of thisinvention can generally be prepared by carrying out the proceduresdisclosed in the Schemes and/or in the Examples below, by substituting areadily available isotopically labeled reagent for a non-isotopicallylabeled reagent.

The invention further provides a pharmaceutical composition (alsoreferred to as pharmaceutical formulation) comprising a compound ofFormula (I) or pharmaceutically acceptable salt thereof and one or moreexcipients (also referred to as carriers and/or diluents in thepharmaceutical arts). The excipients are acceptable in the sense ofbeing compatible with the other ingredients of the formulation and notdeleterious to the recipient thereof (i.e., the patient).

Suitable pharmaceutically acceptable excipients will vary depending uponthe particular dosage form chosen. In addition, suitablepharmaceutically acceptable excipients may be chosen for a particularfunction that they may serve in the composition. For example, certainpharmaceutically acceptable excipients may be chosen for their abilityto facilitate the production of uniform dosage forms. Certainpharmaceutically acceptable excipients may be chosen for their abilityto facilitate the production of stable dosage forms. Certainpharmaceutically acceptable excipients may be chosen for their abilityto facilitate the carrying or transporting of the compound or compoundsof the invention once administered to the patient from one organ, orportion of the body, to another organ, or portion of the body. Certainpharmaceutically acceptable excipients may be chosen for their abilityto enhance patient compliance.

Suitable pharmaceutically acceptable excipients include the followingtypes of excipients: diluents, fillers, binders, disintegrants,lubricants, glidants, granulating agents, coating agents, wettingagents, solvents, co-solvents, suspending agents, emulsifiers,sweeteners, flavoring agents, flavor masking agents, coloring agents,anticaking agents, hemectants, chelating agents, plasticizers, viscosityincreasing agents, antioxidants, preservatives, stabilizers,surfactants, and buffering agents. The skilled artisan will appreciatethat certain pharmaceutically acceptable excipients may serve more thanone function and may serve alternative functions depending on how muchof the excipient is present in the formulation and what otheringredients are present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enablethem to select suitable pharmaceutically acceptable excipients inappropriate amounts for use in the invention. In addition, there are anumber of resources that are available to the skilled artisan whichdescribe pharmaceutically acceptable excipients and may be useful inselecting suitable pharmaceutically acceptable excipients. Examplesinclude Remington's Pharmaceutical Sciences (Mack Publishing Company),The Handbook of Pharmaceutical Additives (Gower Publishing Limited), andThe Handbook of Pharmaceutical Excipients (the American PharmaceuticalAssociation and the Pharmaceutical Press).

The pharmaceutical compositions of the invention are prepared usingtechniques and methods known to those skilled in the art. Some of themethods commonly used in the art are described in Remington'sPharmaceutical Sciences (Mack Publishing Company).

Pharmaceutical compositions may be in unit dose form containing apredetermined amount of active ingredient per unit dose. Such a unit maycontain a therapeutically effective dose of the compound of Formula (I)or salt thereof or a fraction of a therapeutically effective dose suchthat multiple unit dosage forms might be administered at a given time toachieve the desired therapeutically effective dose. Preferred unitdosage formulations are those containing a daily dose or sub-dose, asherein above recited, or an appropriate fraction thereof, of an activeingredient. Furthermore, such pharmaceutical compositions may beprepared by any of the methods well-known in the pharmacy art.

Pharmaceutical compositions may be adapted for administration by anyappropriate route, for example, by oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual, ortransdermal), vaginal, or parenteral (including subcutaneous,intramuscular, intravenous, or intradermal) routes. Such compositionsmay be prepared by any method known in the art of pharmacy, for example,by bringing into association the active ingredient with theexcipient(s).

When adapted for oral administration, pharmaceutical compositions may bein discrete units such as tablets or capsules; powders or granules;solutions or suspensions in aqueous or non-aqueous liquids; edible foamsor whips; oil-in-water liquid emulsions or water-in-oil liquidemulsions. The compound or salt thereof of the invention or thepharmaceutical composition of the invention may also be incorporatedinto a candy, a wafer, and/or tongue tape formulation for administrationas a “quick-dissolve” medicine.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic pharmaceutically acceptable inert carrier such as ethanol,glycerol, water, and the like. Powders or granules are prepared bycomminuting the compound to a suitable fine size and mixing with asimilarly comminuted pharmaceutical carrier such as an ediblecarbohydrate, as, for example, starch or mannitol. Flavoring,preservative, dispersing, and coloring agents can also be present.

Capsules are made by preparing a powder mixture, as described above, andfilling formed gelatin or non-gelatinous sheaths. Glidants andlubricants such as colloidal silica, talc, magnesium stearate, calciumstearate, solid polyethylene glycol can be added to the powder mixturebefore the filling operation. A disintegrating or solubilizing agentsuch as agar-agar, calcium carbonate, or sodium carbonate can also beadded to improve the availability of the medicine when the capsule isingested.

Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents, and coloring agents can also be incorporated intothe mixture. Suitable binders include starch, gelatin, natural sugars,such as glucose or beta-lactose, corn sweeteners, natural and syntheticgums such as acacia, tragacanth, sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes, and the like.Lubricants used in these dosage forms include sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate, sodiumchloride, and the like. Disintegrators include, without limitation,starch, methylcellulose, agar, bentonite, xanthan gum, and the like.

Tablets are formulated, for example, by preparing a powder mixture,granulating or slugging, adding a lubricant and disintegrant, andpressing into tablets. A powder mixture is prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove, and optionally, with a binder such as carboxymethylcellulose, andaliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant suchas paraffin, a resorption accelerator such as a quaternary salt, and/oran absorption agent such as bentonite, kaolin, or dicalcium phosphate.The powder mixture can be granulated by wetting a binder such as syrup,starch paste, acadia mucilage, or solutions of cellulosic or polymericmaterials and forcing through a screen. As an alternative togranulating, the powder mixture can be run through the tablet machineand the result is imperfectly formed slugs broken into granules. Thegranules can be lubricated to prevent sticking to the tablet formingdies by means of the addition of stearic acid, a stearate salt, talc, ormineral oil. The lubricated mixture is then compressed into tablets. Thecompound or salt of the present invention can also be combined with afree-flowing inert carrier and compressed into tablets directly withoutgoing through the granulating or slugging steps. A clear opaqueprotective coating consisting of a sealing coat of shellac, a coating ofsugar, or polymeric material, and a polish coating of wax can beprovided. Dyestuffs can be added to these coatings to distinguishdifferent dosages.

Oral fluids such as solutions, syrups, and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of active ingredient. Syrups can be prepared by dissolving thecompound or salt thereof of the invention in a suitably flavouredaqueous solution, while elixirs are prepared through the use of anon-toxic alcoholic vehicle. Suspensions can be formulated by dispersingthe compound or salt of the invention in a non-toxic vehicle.Solubilizers and emulsifiers, such as ethoxylated isostearyl alcoholsand polyoxyethylene sorbitol ethers, preservatives, flavor additivessuch as peppermint oil, natural sweeteners, saccharin, or otherartificial sweeteners, and the like, can also be added.

Where appropriate, dosage unit formulations for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as, for example, by coating or embedding particulatematerial in polymers, wax, or the like.

In the present invention, tablets and capsules are preferred fordelivery of the pharmaceutical composition.

In accordance with another aspect of the invention there is provided aprocess for the preparation of a pharmaceutical composition comprisingmixing (or admixing) a compound of Formula (I) or salt thereof with atleast one excipient.

The present invention also provides a method of treatment in a mammal,especially a human. The compounds and compositions of the invention areused to treat cellular proliferation diseases. Disease states which canbe treated by the methods and compositions provided herein include, butare not limited to, cancer (further discussed below), autoimmunedisease, fungal disorders, arthritis, graft rejection, inflammatorybowel disease, proliferation induced after medical procedures,including, but not limited to, surgery, angioplasty, and the like. It isappreciated that in some cases the cells may not be in a hyper or hypoproliferation state (abnormal state) and still requires treatment. Forexample, during wound healing, the cells may be proliferating“normally”, but proliferation enhancement may be desired. Thus, in oneembodiment, the invention herein includes application to cells orindividuals afflicted or impending affliction with any one of thesedisorders or states.

The compositions and methods provided herein are particularly deemeduseful for the treatment of cancer including tumors such as prostate,breast, brain, skin, cervical carcinomas, testicular carcinomas, etc.They are particularly useful in treating metastatic or malignant tumors.More particularly, cancers that may be treated by the compositions andmethods of the invention include, but are not limited to tumor typessuch as astrocytic, breast, cervical, colorectal, endometrial,esophageal, gastric, head and neck, hepatocellular, laryngeal, lung,oral, ovarian, prostate and thyroid carcinomas and sarcomas. Morespecifically, these compounds can be used to treat: Cardiac: sarcoma(angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma,rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma(squamous cell, undifferentiated small cell, undifferentiated largecell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchialadenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma;Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma,leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma,leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma,glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel(adenocarcinoma, lymphoma, carcinoid tumors, Karposi's sarcoma,leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel(adenocarcinoma, tubular adenoma, villous adenoma, hamartoma,leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor(nephroblastoma), lymphoma, leukemia), bladder and urethra (squamouscell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonalcarcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cellcarcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver:hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma,angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gallbladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone:osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibroushistiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma(reticulum cell sarcoma), multiple myeloma, malignant giant cell tumorchordoma, osteochronfroma (osteocartilaginous exostoses), benignchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma andgiant cell tumors; Nervous system: skull (osteoma, hemangioma,granuloma, xanthoma, osteitis deformans), meninges (meningioma,meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform,oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological:uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumorcervical dysplasia), ovaries (ovarian carcinoma (serouscystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiedcarcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors,dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma),vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma(embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic:blood (myeloid leukemia (acute and chronic), acute lymphoblasticleukemia, chronic lymphocytic leukemia, myeloproliferative diseases,multiple myeloma, myelodysplastic syndrome), Hodgkin's disease,non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma,basal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma, molesdysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis;and Adrenal glands: neuroblastoma. Thus, the term “cancerous cell” asprovided herein, includes a cell afflicted by any one or related of theabove identified conditions.

The instant compounds can be combined with or co-administered with othertherapeutic agents, particularly agents that may enhance the activity ortime of disposition of the compounds. Combination therapies according tothe invention comprise the administration of at least one compound ofthe invention and the use of at least one other treatment method. In oneembodiment, combination therapies according to the invention comprisethe administration of at least one compound of the invention andsurgical therapy. In one embodiment, combination therapies according tothe invention comprise the administration of at least one compound ofthe invention and radiotherapy. In one embodiment, combination therapiesaccording to the invention comprise the administration of at least onecompound of the invention and at least one supportive care agent (e.g.,at least one anti-emetic agent). In one embodiment, combinationtherapies according to the present invention comprise the administrationof at least one compound of the invention and at least one otherchemotherapeutic agent. In one particular embodiment, the inventioncomprises the administration of at least one compound of the inventionand at least one anti-neoplastic agent. In yet another embodiment, theinvention comprises a therapeutic regimen where the EZH2 inhibitors ofthis disclosure are not in and of themselves active or significantlyactive, but when combined with another therapy, which may or may not beactive as a standalone therapy, the combination provides a usefultherapeutic outcome.

By the term “co-administering” and derivatives thereof as used hereinrefers to either simultaneous administration or any manner of separatesequential administration of an EZH2 inhibiting compound, as describedherein, and a further active ingredient or ingredients, known to beuseful in the treatment of cancer, including chemotherapy and radiationtreatment. The term further active ingredient or ingredients, as usedherein, includes any compound or therapeutic agent known to or thatdemonstrates advantageous properties when administered to a patient inneed of treatment for cancer. Preferably, if the administration is notsimultaneous, the compounds are administered in a close time proximityto each other. Furthermore, it does not matter if the compounds areadministered in the same dosage form, e.g. one compound may beadministered topically and another compound may be administered orally.

Typically, any anti-neoplastic agent that has activity versus asusceptible tumor being treated may be co-administered in the treatmentof specified cancers in the present invention. Examples of such agentscan be found in Cancer Principles and Practice of Oncology by V. T.Devita and S. Hellman (editors), 6^(th) edition (Feb. 15, 2001),Lippincott Williams & Wilkins Publishers. A person of ordinary skill inthe art would be able to discern which combinations of agents would beuseful based on the particular characteristics of the drugs and thecancer involved. Typical anti-neoplastic agents useful in the presentinvention include, but are not limited to, anti-microtubule agents suchas diterpenoids and vinca alkaloids; platinum coordination complexes;alkylating agents such as nitrogen mustards, oxazaphosphorines,alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents such asanthracyclins, actinomycins and bleomycins; topoisomerase II inhibitorssuch as epipodophyllotoxins; antimetabolites such as purine andpyrimidine analogues and anti-folate compounds; topoisomerase Iinhibitors such as camptothecins; hormones and hormonal analogues; DNAmethyltransferase inhibitors such as azacitidine and decitabine; signaltransduction pathway inhibitors; non-receptor tyrosine kinaseangiogenesis inhibitors; immunotherapeutic agents; proapoptotic agents;and cell cycle signaling inhibitors.

Typically, any chemotherapeutic agent that has activity against asusceptible neoplasm being treated may be utilized in combination withthe compounds the invention, provided that the particular agent isclinically compatible with therapy employing a compound of theinvention. Typical anti-neoplastic agents useful in the presentinvention include, but are not limited to: alkylating agents,anti-metabolites, antitumor antibiotics, antimitotic agents, nucleosideanalogues, topoisomerase I and II inhibitors, hormones and hormonalanalogues; retinoids, histone deacetylase inhibitors; signaltransduction pathway inhibitors including inhibitors of cell growth orgrowth factor function, angiogenesis inhibitors, and serine/threonine orother kinase inhibitors; cyclin dependent kinase inhibitors; antisensetherapies and immunotherapeutic agents, including monoclonals, vaccinesor other biological agents.

Nucleoside analogues are those compounds which are converted todeoxynucleotide triphosphates and incorporated into replicating DNA inplace of cytosine.

DNA methyltransferases become covalently bound to the modified basesresulting in an inactive enzyme and reduced DNA methylation. Examples ofnucleoside analogues include azacitidine and decitabine which are usedfor the treatment of myelodysplastic disorder. Histone deacetylase(HDAC) inhibitors include vorinostat, for the treatment of cutaneousT-cell lymphoma. HDACs modify chromatin through the deacetylation ofhistones. In addition, they have a variety of substrates includingnumerous transcription factors and signaling molecules. Other HDACinhibitors are in development.

Signal transduction pathway inhibitors are those inhibitors which blockor inhibit a chemical process which evokes an intracellular change. Asused herein this change is cell proliferation or differentiation orsurvival. Signal transduction pathway inhibitors useful in the presentinvention include, but are not limited to, inhibitors of receptortyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domainblockers, serine/threonine kinases, phosphatidyl inositol-3-OH kinases,myoinositol signaling, and Ras oncogenes. Signal transduction pathwayinhibitors may be employed in combination with the compounds of theinvention in the compositions and methods described above.

Receptor kinase angiogenesis inhibitors may also find use in the presentinvention. Inhibitors of angiogenesis related to VEGFR and TIE-2 arediscussed above in regard to signal transduction inhibitors (both arereceptor tyrosine kinases). Other inhibitors may be used in combinationwith the compounds of the invention. For example, anti-VEGF antibodies,which do not recognize VEGFR (the receptor tyrosine kinase), but bind tothe ligand; small molecule inhibitors of integrin (alpha, beta₃) thatinhibit angiogenesis; endostatin and angiostatin (non-RTK) may alsoprove useful in combination with the compounds of the invention. Oneexample of a VEGFR antibody is bevacizumab (AVASTIN®)).

Several inhibitors of growth factor receptors are under development andinclude ligand antagonists, antibodies, tyrosine kinase inhibitors,anti-sense oligonucleotides and aptamers. Any of these growth factorreceptor inhibitors may be employed in combination with the compounds ofthe invention in any of the compositions and methods/uses describedherein. Trastuzumab (Herceptin®) is an example of an anti-erbB2 antibodyinhibitor of growth factor function. One example of an anti-erbB1antibody inhibitor of growth factor function is cetuximab (Erbitux™,C225). Bevacizumab (Avastin®) is an example of a monoclonal antibodydirected against VEGFR. Examples of small molecule inhibitors ofepidermal growth factor receptors include but are not limited tolapatinib (Tykerb) and erlotinib (TARCEVA®). Imatinib mesylate (GLEEVEC)is one example of a PDGFR inhibitor. Examples of VEGFR inhibitorsinclude pazopanib (Votrient®), ZD6474, AZD2171, PTK787, sunitinib andsorafenib.

Anti-microtubule or anti-mitotic agents are phase specific agents activeagainst the microtubules of tumor cells during M or the mitosis phase ofthe cell cycle. Examples of anti-microtubule agents include, but are notlimited to, diterpenoids and vinca alkaloids.

Diterpenoids, which are derived from natural sources, are phase specificanti-cancer agents that operate at the G₂/M phases of the cell cycle. Itis believed that the diterpenoids stabilize the β-tubulin subunit of themicrotubules, by binding with this protein. Disassembly of the proteinappears then to be inhibited with mitosis being arrested and cell deathfollowing. Examples of diterpenoids include, but are not limited to,paclitaxel and its analog docetaxel.

Paclitaxel, 5β,20-epoxy-1,2α,4,7β,10β,13α-hexa-hydroxytax-11-en-9-one4,10-diacetate 2-benzoate 13-ester with(2R,3S)—N-benzoyl-3-phenylisoserine; is a natural diterpene productisolated from the Pacific yew tree Taxus brevifolia and is commerciallyavailable as an injectable solution TAXOL®. It is a member of the taxanefamily of terpenes. It was first isolated in 1971 by Wani et al. J. Am.Chem, Soc., 93:2325 (1971), who characterized its structure by chemicaland X-ray crystallographic methods. One mechanism for its activityrelates to paclitaxel's capacity to bind tubulin, thereby inhibitingcancer cell growth. Schiff et al., Proc. Natl, Acad, Sci. USA,77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar,J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis andanticancer activity of some paclitaxel derivatives see: D. G. I.Kingston et al., Studies in Organic Chemistry vol. 26, entitled “Newtrends in Natural Products Chemistry 1986”, Attaur-Rahman, P. W. LeQuesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.

Paclitaxel has been approved for clinical use in the treatment ofrefractory ovarian cancer in the United States (Markman et al., YaR^(c)Journal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann. Int.Med., 111:273,1989) and for the treatment of breast cancer (Holmes etal., J. Nat. Cancer Inst., 83:1797,1991.). It is a potential candidatefor treatment of neoplasms in the skin (Einzig et. al., Proc. Am. Soc.Clin. Oncol., 20:46) and head and neck carcinomas (Forastire et. al.,Sem. Oncol., 20:56, 1990).

The compound also shows potential for the treatment of polycystic kidneydisease (Woo et. al., Nature, 368:750. 1994), lung cancer and malaria.Treatment of patients with paclitaxel results in bone marrow suppression(multiple cell lineages, Ignoff, R. J. et. al, Cancer ChemotherapyPocket Guide,. 1998) related to the duration of dosing above a thresholdconcentration (50 nM) (Kearns, C. M. et. al., Seminars in Oncology, 3(6)p.16-23, 1995).

Docetaxel, (2R,3S)- N-carboxy-3-phenylisoserine N-tert-butyl ester,13-ester with 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one4-acetate 2-benzoate, trihydrate; is commercially available as aninjectable solution as TAXOTERE®. Docetaxel is indicated for thetreatment of breast cancer. Docetaxel is a semisynthetic derivative ofpaclitaxel q.v., prepared using a natural precursor,10-deacetyl-baccatin III, extracted from the needle of the European Yewtree. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived fromthe periwinkR^(c) plant. Vinca alkaloids act at the M phase (mitosis) ofthe cell cycle by binding specifically to tubulin. Consequently, thebound tubulin molecule is unable to polymerize into microtubules.Mitosis is believed to be arrested in metaphase with cell deathfollowing. Examples of vinca alkaloids include, but are not limited to,vinblastine, vincristine, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available asVELBAN® as an injectable solution. Although, it has possible indicationas a second line therapy of various solid tumors, it is primarilyindicated in the treatment of testicular cancer and various lymphomasincluding Hodgkin's Disease; and lymphocytic and histiocytic lymphomas.Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commerciallyavailable as ONCOVIN® as an injectable solution. Vincristine isindicated for the treatment of acute leukemias and has also found use intreatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas.Alopecia and neurologic effects are the most common side effect ofvincristine and to a lesser extent myelosupression and gastrointestinalmucositis effects occur.

Vinorelbine, 3′,4′-didehydro -4′-deoxy-C′-norvincaleukoblastine[R-(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)], commerciallyavailable as an injectable solution of vinorelbine tartrate(NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine isindicated as a single agent or in combination with otherchemotherapeutic agents, such as cisplatin, in the treatment of varioussolid tumors, particularly non-small cell lung, advanced breast, andhormone refractory prostate cancers. Myelosuppression is the most commondose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-canceragents, which are interactive with DNA. The platinum complexes entertumor cells, undergo aquation and form intra- and interstrand crosslinkswith DNA causing adverse biological effects to the tumor. Examples ofplatinum coordination complexes include, but are not limited to,cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available asPLATINOL® as an injectable solution. Cisplatin is primarily indicated inthe treatment of metastatic testicular and ovarian cancer and advancedbladder cancer. The primary dose limiting side effects of cisplatin arenephrotoxicity, which may be controlled by hydration and diuresis, andototoxicity.

Carboplatin, platinum, diammine[1,1-cyclobutane-dicarboxylate(2-)-O,O′], is commercially available asPARAPLATIN® as an injectable solution. Carboplatin is primarilyindicated in the first and second line treatment of advanced ovariancarcinoma. Bone marrow suppression is the dose limiting toxicity ofcarboplatin.

Alkylating agents are non-phase anti-cancer specific agents and strongelectrophiles. Typically, alkylating agents form covalent linkages, byalkylation, to DNA through nucleophilic moieties of the DNA moleculesuch as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazolegroups. Such alkylation disrupts nucleic acid function leading to celldeath. Examples of alkylating agents include, but are not limited to,nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil;alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; andtriazenes such as dacarbazine.

Cyclophosphamide,2-[bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxidemonohydrate, is commercially available as an injectable solution ortablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent orin combination with other chemotherapeutic agents, in the treatment ofmalignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea,vomiting and leukopenia are the most common dose limiting side effectsof cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commerciallyavailable as an injectable solution or tablets as ALKERAN®. Melphalan isindicated for the palliative treatment of multiple myeloma andnon-resectable epithelial carcinoma of the ovary. Bone marrowsuppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, iscommercially available as LEUKERAN® tablets. Chlorambucil is indicatedfor the palliative treatment of chronic lymphatic leukemia, andmalignant lymphomas such as lymphosarcoma, giant follicular lymphoma,and Hodgkin's disease. Bone marrow suppression is the most common doselimiting side effect of chlorambucil.

Busulfan, 1,4-butanediol dimethanesulfonate, is commercially availableas MYLERAN® TABLETS. Busulfan is indicated for the palliative treatmentof chronic myelogenous leukemia. Bone marrow suppression is the mostcommon dose limiting side effects of busulfan.

Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commerciallyavailable as single vials of lyophilized material as BiCNU®. Carmustineis indicated for the palliative treatment as a single agent or incombination with other agents for brain tumors, multiple myeloma,Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppressionis the most common dose limiting side effects of carmustine.

Dacarbazine, 5-(3,3-dimethyl-l-triazeno)-imidazoR^(c)-4-carboxamide, iscommercially available as single vials of material as DTIC-Dome®.Dacarbazine is indicated for the treatment of metastatic malignantmelanoma and in combination with other agents for the second linetreatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are themost common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplastics are non-phase specific agents, which bind orintercalate with DNA. Typically, such action results in stable DNAcomplexes or strand breakage, which disrupts ordinary function of thenucleic acids leading to cell death. Examples of antibioticanti-neoplastic agents include, but are not limited to, actinomycinssuch as dactinomycin, anthrocyclins such as daunorubicin anddoxorubicin; and bleomycins.

Dactinomycin, also known as Actinomycin D, is commercially available ininjectable form as COSMEGEN®. Dactinomycin is indicated for thetreatment of Wilm's tumor and rhabdomyosarcoma. Nausea, vomiting, andanorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin,(8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12naphthacenedione hydrochloride, is commercially available as a liposomalinjectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.Daunorubicin is indicated for remission induction in the treatment ofacute nonlymphocytic leukemia and advanced HIV associated Kaposi'ssarcoma. Myelosuppression is the most common dose limiting side effectof daunorubicin.

Doxorubicin, (8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-8-glycoloyl,7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12 naphthacenedionehydrochloride, is commercially available as an injectable form as RUBEX®or ADRIAMYCIN RDF®. Doxorubicin is primarily indicated for the treatmentof acute lymphoblastic leukemia and acute myeloblastic leukemia, but isalso a useful component in the treatment of some solid tumors andlymphomas. Myelosuppression is the most common dose limiting side effectof doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated froma strain of Streptomyces verticillus, is commercially available asBLENOXANE®. Bleomycin is indicated as a palliative treatment, as asingle agent or in combination with other agents, of squamous cellcarcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneoustoxicities are the most common dose limiting side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to,epipodophyllotoxins.

Epipodophyllotoxins are phase specific anti-neoplastic agents derivedfrom the mandrake plant. Epipodophyllotoxins typically affect cells inthe S and G₂ phases of the cell cycle by forming a ternary complex withtopoisomerase II and DNA causing DNA strand breaks. The strand breaksaccumulate and cell death follows. Examples of epipodophyllotoxinsinclude, but are not limited to, etoposide and teniposide.

Etoposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially availableas an injectable solution or capsules as VePESID® and is commonly knownas VP-16. Etoposide is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of testicular andnon-small cell lung cancers. Myelosuppression is the most common sideeffect of etoposide. The incidence of leukopenialeukopenia tends to bemore severe than thrombocytopenia.

Teniposide, 4′-demethyl-epipodophyllotoxin9[4,6-0-(R)-thenylidene-13-D-glucopyranoside], is commercially availableas an injectable solution as VUMON® and is commonly known as VM-26.Teniposide is indicated as a single agent or in combination with otherchemotherapy agents in the treatment of acute leukemia in children.Myelosuppression is the most common dose limiting side effect ofteniposide. Teniposide can induce both leukopenialeukopenia andthrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplasticagents that act at S phase (DNA synthesis) of the cell cycle byinhibiting DNA synthesis or by inhibiting purine or pyrimidine basesynthesis and thereby limiting DNA synthesis. Consequently, S phase doesnot proceed and cell death follows. Examples of antimetaboliteanti-neoplastic agents include, but are not limited to, fluorouracil,methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine.

5-fluorouracil, 5-fluoro-2,4- (1H,3H) pyrimidinedione, is commerciallyavailable as fluorouracil. Administration of 5-fluorouracil leads toinhibition of thymidylate synthesis and is also incorporated into bothRNA and DNA. The result typically is cell death. 5-fluorouracil isindicated as a single agent or in combination with other chemotherapyagents in the treatment of carcinomas of the breast, colon, rectum,stomach and pancreas. Myelosuppression and mucositis are dose limitingside effects of 5-fluorouracil. Other fluoropyrimidine analogs include5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridinemonophosphate.

Cytarabine, 4-amino-1-13-D-arabinofuranosyl-2 (1H)-pyrimidinone, iscommercially available as CYTOSAR-U® and is commonly known as Ara-C. Itis believed that cytarabine exhibits cell phase specificity at S-phaseby inhibiting DNA chain elongation by terminal incorporation ofcytarabine into the growing DNA chain. Cytarabine is indicated as asingle agent or in combination with other chemotherapy agents in thetreatment of acute leukemia. Other cytidine analogs include5-azacytidine and 2′,2′-difluorodeoxycytidine (gemcitabine). Cytarabineinduces leukopenialeukopenia, thrombocytopenia, and mucositis.

Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, iscommercially available as PURINETHOL®. Mercaptopurine exhibits cellphase specificity at S-phase by inhibiting DNA synthesis by an as of yetunspecified mechanism. Mercaptopurine is indicated as a single agent orin combination with other chemotherapy agents in the treatment of acuteleukemia. Myelosuppression and gastrointestinal mucositis are expectedside effects of mercaptopurine at high doses. A useful mercaptopurineanalog is azathioprine.

Thioguanine, 2-amino-1,7-dihydro-6H-purine-6-thione, is commerciallyavailable as TABLOID®. Thioguanine exhibits cell phase specificity atS-phase by inhibiting DNA synthesis by an as of yet unspecifiedmechanism. Thioguanine is indicated as a single agent or in combinationwith other chemotherapy agents in the treatment of acute leukemia.Myelosuppression, including leukopenialeukopenia, thrombocytopenia, andanemia, is the most common dose limiting side effect of thioguanineadministration. However, gastrointestinal side effects occur and can bedose limiting. Other purine analogs include pentostatin,erythrohydroxynonyladenine, fludarabine phosphate, and cladribine.

Gemcitabine, 2′-deoxy-2′, 2′-difluorocytidine monohydrochloride((3-isomer), is commercially available as GEMZAR®. Gemcitabine exhibitscell phase specificity at S-phase and by blocking progression of cellsthrough the Gl/S boundary. Gemcitabine is indicated in combination withcisplatin in the treatment of locally advanced non-small cell lungcancer and alone in the treatment of locally advanced pancreatic cancer.Myelosuppression, including leukopenialeukopenia, thrombocytopenia, andanemia, is the most common dose limiting side effect of gemcitabineadministration.

Methotrexate,N-[4[[(2,4-diamino-6-pteridinyl)methyl]methylamino]benzoyl]-L-glutamicacid, is commercially available as methotrexate sodium. Methotrexateexhibits cell phase effects specifically at S-phase by inhibiting DNAsynthesis, repair and/or replication through the inhibition ofdyhydrofolic acid reductase which is required for synthesis of purinenucleotides and thymidylate. Methotrexate is indicated as a single agentor in combination with other chemotherapy agents in the treatment ofchoriocarcinoma, meningeal leukemia, non-Hodgkin's lymphoma, andcarcinomas of the breast, head, neck, ovary and bladder.Myelosuppression (leukopenia, thrombocytopenia, and anemia) andmucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives areavailable or under development as Topoisomerase I inhibitors.Camptothecins cytotoxic activity is believed to be related to itsTopoisomerase I inhibitory activity. Examples of camptothecins include,but are not limited to irinotecan, topotecan, and the various opticalforms of7-(4-methylpiperazino-methylene)-10,11-ethylenedioxy-20-camptothecindescribed below.

Irinotecan HCl, (4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride, is commercially available as the injectable solutionCAMPTOSAR®.

Irinotecan is a derivative of camptothecin which binds, along with itsactive metabolite SN-38, to the topoisomerase I-DNA complex. It isbelieved that cytotoxicity occurs as a result of irreparable doublestrand breaks caused by interaction of the topoisomerase I:DNA:irintecanor SN-38 ternary complex with replication enzymes. Irinotecan isindicated for treatment of metastatic cancer of the colon or rectum. Thedose limiting side effects of irinotecan HC1 are myelosuppression,including neutropenia, and GI effects, including diarrhea.

Topotecan HCl,(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′,6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dionemonohydrochloride, is commercially available as the injectable solutionHYCAMTIN®. Topotecan is a derivative of camptothecin which binds to thetopoisomerase I-DNA complex and prevents religation of singles strandbreaks caused by Topoisomerase I in response to torsional strain of theDNA molecule. Topotecan is indicated for second line treatment ofmetastatic carcinoma of the ovary and small cell lung cancer. The doselimiting side effect of topotecan HCl is myelosuppression, primarilyneutropenia.

Pharmaceutical compositions may be presented in unit dose formscontaining a predetermined amount of active ingredient per unit dose.Such a unit may contain, for example, 0.5 mg to 1 g, preferably 1 mg to700 mg, more preferably 5 mg to 100 mg of a compound of the Formula (I),depending on the condition being treated, the route of administrationand the age, weight and condition of the patient, or pharmaceuticalcompositions may be presented in unit dose forms containing apredetermined amount of active ingredient per unit dose. Preferred unitdosage compositions are those containing a daily dose or sub-dose, asherein above recited, or an appropriate fraction thereof, of an activeingredient. Furthermore, such pharmaceutical compositions may beprepared by any of the methods well known in the pharmacy art.

Pharmaceutical compositions may be adapted for administration by anyappropriate route, for example by the oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual ortransdermal), vaginal or parenteral (including subcutaneous,intramuscular, intravenous or intradermal) route. Such compositions maybe prepared by any method known in the art of pharmacy, for example bybringing into association a compound of formal (I) with the carrier(s)or excipient(s).

Pharmaceutical compositions adapted for oral administration may bepresented as discrete units such as capsules or tablets; powders orgranules; solutions or suspensions in aqueous or non-aqueous liquids;edible foams or whips; or oil-in-water liquid emulsions or water-in-oilliquid emulsions.

Capsules are made by preparing a powder mixture, as described above, andfilling formed gelatin sheaths. Glidants and lubricants such ascolloidal silica, talc, magnesium stearate, calcium stearate or solidpolyethylene glycol can be added to the powder mixture before thefilling operation. A disintegrating or solubilizing agent such asagar-agar, calcium carbonate or sodium carbonate can also be added toimprove the availability of the medicament when the capsule is ingested.

Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents and coloring agents can also be incorporated intothe mixture. Suitable binders include starch, gelatin, natural sugarssuch as glucose or beta-lactose, corn sweeteners, natural and syntheticgums such as acacia, tragacanth or sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes and the like.Lubricants used in these dosage forms include sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate, sodiumchloride and the like. Disintegrators include, without limitation,starch, methyl cellulose, agar, bentonite, xanthan gum and the like.Tablets are formulated, for example, by preparing a powder mixture,granulating or slugging, adding a lubricant and disintegrant andpressing into tablets. A powder mixture is prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove, and optionally, with a binder such as carboxymethylcellulose, analiginate, gelatin, or polyvinyl pyrrolidone, a solution retardant suchas paraffin, a resorption accelerator such as a quaternary salt and/oran absorption agent such as bentonite, kaolin or dicalcium phosphate.The powder mixture can be granulated by tablet forming dies by means ofthe addition of stearic acid, a stearate salt, talc or mineral oil. Thelubricated mixture is then compressed into tablets. The compounds of thepresent invention can also be combined with a free flowing inert carrierand compressed into tablets directly without going through thegranulating or slugging steps. A clear or opaque protective coatingconsisting of a sealing coat of shellac, a coating of sugar or polymericmaterial and a polish coating of wax can be provided. Dyestuffs can beadded to these coatings to distinguish different unit dosages.

Oral fluids such as solution, syrups and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of a compound of Formula (I). Syrups can be prepared bydissolving the compound in a suitably flavored aqueous solution, whileelixirs are prepared through the use of a non-toxic alcoholic vehicle.Suspensions can be formulated by dispersing the compound in a non-toxicvehicle. Solubilizers and emulsifiers such as ethoxylated isostearylalcohols and polyoxy ethylene sorbitol ethers, preservatives, flavoradditive such as peppermint oil or natural sweeteners or saccharin orother artificial sweeteners, and the like can also be added.

Where appropriate, dosage unit pharmaceutical compositions for oraladministration can be microencapsulated. The formulation can also beprepared to prolong or sustain the release as for example by coating orembedding particulate material in polymers, wax or the like.

Pharmaceutical compositions adapted for rectal administration may bepresented as suppositories or as enemas.

Pharmaceutical compositions adapted for vaginal administration may bepresented as pessaries, tampons, creams, gels, pastes, foams or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats and solutes which renderthe composition isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The pharmaceutical compositions may bepresented in unit-dose or multi-dose containers, for example sealedampoules and vials, and may be stored in a freeze-dried (lyophilized)condition requiring only the addition of the sterile liquid carrier, forexample water for injections, immediately prior to use. Extemporaneousinjection solutions and suspensions may be prepared from sterilepowders, granules and tablets.

It should be understood that in addition to the ingredients particularlymentioned above, the pharmaceutical compositions may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

A therapeutically effective amount of a compound of the presentinvention will depend upon a number of factors including, for example,the age and weight of the intended recipient, the precise conditionrequiring treatment and its severity, the nature of the formulation, andthe route of administration, and will ultimately be at the discretion ofthe attendant prescribing the medication. However, an effective amountof a compound of Formula (I) for the treatment of anemia will generallybe in the range of 0.001 to 100 mg/kg body weight of recipient per day,suitably in the range of 0.01 to 10 mg/kg body weight per day. For a 70kg adult mammal, the actual amount per day would suitably be from 7 to700 mg and this amount may be given in a single dose per day or in anumber (such as two, three, four, five or six) of sub-doses per day suchthat the total daily dose is the same. An effective amount of a salt orsolvate, etc., may be determined as a proportion of the effective amountof the compound of Formula (I) per se. It is envisaged that similardosages would be appropriate for treatment of the other conditionsreferred to above.

DEFINITIONS

Terms are used within their accepted meanings. The following definitionsare meant to clarify, but not limit, the terms defined.

As used herein, the term “alkyl” represents a saturated, straight orbranched hydrocarbon moiety having the specified number of carbon atoms.The term “(C₁-C₆)alkyl” refers to an alkyl moiety containing from 1 to 6carbon atoms. Exemplary alkyls include, but are not limited to methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl,and hexyl.

When the term “alkyl” is used in combination with other substituentgroups, such as “halo(C₁-C₄)alkyl”, “hydroxy(C₁-C₄)alkyl” or“aryl(C₁-C₄)alkyl-”, the term “alkyl” is intended to encompass adivalent straight or branched-chain hydrocarbon radical, wherein thepoint of attachment is through the alkyl moiety. The term“halo(C₁-C₄)alkyl” is intended to mean a radical having one or morehalogen atoms, which may be the same or different, at one or more carbonatoms of an alkyl moiety containing from 1 to 4 carbon atoms, which is astraight or branched-chain carbon radical. Examples of“halo(C₁-C₄)alkyl” groups useful in the present invention include, butare not limited to, —CF₃ (trifluoromethyl), —CC1₃ (trichloromethyl),1,1-difluoroethyl, 2,2,2-trifluoroethyl, and hexafluoroisopropyl.Examples of “aryl(C₁-C₄)alkyl-” groups useful in the present inventioninclude, but are not limited to, benzyl (phenylmethyl), 1-methylbenzyl(1-phenylethyl), 1,1-dimethylbenzyl (1-phenylisopropyl), and phenethyl(2-phenylethyl). Examples of “hydroxy(C₁-C₄)alkyl” groups useful in thepresent invention include, but are not limited to, hydroxymethyl,hydroxyethyl, and hydroxyisopropyl.

“Alkoxy” refers to a group containing an alkyl radical, definedhereinabove, attached through an oxygen linking atom. The term“(C₁-C₄)alkoxy” refers to a straight- or branched-chain hydrocarbonradical having at least 1 and up to 4 carbon atoms attached through anoxygen linking atom. Exemplary “(C₁-C₄)alkoxy” groups useful in thepresent invention include, but are not limited to, methoxy, ethoxy,n-propoxy, isopropoxy, n-butoxy, s-butoxy, isobutoxy, and t-butoxy.

When “cycloalkyl” is used it refers to a non-aromatic, saturated, cyclichydrocarbon ring containing the specified number of carbon atoms. So,for example, the term “(C₃-C₈)cycloalkyl” refers to a non-aromaticcyclic hydrocarbon ring having from three to eight carbon atoms.Exemplary “(C₃-C₈)cycloalkyl” groups useful in the present inventioninclude, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl.

As used herein, the term “cycloalkyloxy-” refers to a group containing acycloalkyl radical, defined hereinabove, attached through an oxygenlinking atom. Exemplary “(C3-C8)cycloalkyloxy-” groups useful in thepresent invention include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy,cyclohexyloxy, cycloheptyloxy, and cyclooctyloxy.

As used herein, the term “bicycloalkyl” refers to a saturated, bridged,fused, or spiro, bicyclic hydrocarbon ring system containing thespecified number of carbon atoms. Exemplary “(C₆-C₁₀)bicycloalkyl”groups include, but are not limited to bicyclo[2.1.1]hexyl,bicyclo[2.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl,bicyclo[3.2.2]nonyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2]decyl,bicyclo[4.3.1]decyl, bicyclo[2.2.0]hexyl, bicyclo[3.1.0]hexyl,bicyclo[3.2.0]heptyl, bicyclo[4.1.0]heptyl, octahydropentalenyl,bicyclo[4.2.0]octyl, decahydronaphthalenyl, spiro[3.3]heptyl,spiro[2.4]heptyl, spiro[3.4]octyl, spiro[2.5]octyl, spiro[4.4]nonyl,spiro[3.5]nonyl, and spiro[4.5]decyl.

The terms “halogen” and “halo” represent chloro, fluoro, bromo, or iodosubstituents. “Hydroxy” or “hydroxyl” is intended to mean the radical—OH.

“Heterocycloalkyl” represents a group or moiety comprising anon-aromatic, monovalent monocyclic or bicyclic radical, which issaturated or partially unsaturated, containing 3 to 10 ring atoms, whichincludes 1 to 3 heteroatoms independently selected from nitrogen, oxygenand sulfur, including N-oxides, sulfur oxides, and dioxides.Illustrative examples of heterocycloalkyls useful in the presentinvention include, but are not limited to, aziridinyl, azetidinyl,pyrrolidinyl, pyrazolidinyl, pyrazolinyl, imidazolidinyl, imidazolinyl,oxazolinyl, thiazolinyl, tetrahydrofuranyl, dihydrofuranyl,1,3-dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl,tetrahydropyranyl, dihydropyranyl, 1,3-dioxanyl, 1,4-dioxanyl,1,3-oxathiolanyl, 1,3-oxathianyl, 1,3-dithianyl, 1,4-dithianyl,hexahydro-1H-1,4-diazepinyl, azabicylo[3.2.1]octyl,azabicylo[3.3.1]nonyl, azabicylo[4.3.0]nonyl, oxabicylo[2.2.1]heptyl,1,1-dioxidotetrahydro-2H-thiopyranyl, and 1,5,9-triazacyclododecyl.

As used herein, the term “heterocycloalkyloxy-” refers to a groupcontaining a heterocycloalkyl radical, defined hereinabove, attachedthrough an oxygen linking atom. Illustrative examples ofheterocycloalkyloxy groups useful in the present invention include, butare not limited to, aziridinyloxy, azetidinyloxy, pyrrolidinyloxy,pyrazolidinyloxy, pyrazolinyloxy, imidazolidinyloxy, imidazolinyloxy,oxazolinyloxy, thiazolinyloxy, tetrahydrofuranyloxy, dihydrofuranyloxy,1,3-dioxolanyloxy, piperidinyloxy, piperazinyloxy, morpholinyloxy,thiomorpholinyloxy, tetrahydropyranyloxy, dihydropyranyloxy,1,3-dioxanyloxy, 1,4-dioxanyloxy, 1,3-oxathiolanyloxy,1,3-oxathianyloxy, 1,3-dithianyloxy, hexahydro-1H-1,4-diazepinyloxy,azabicylo[3.2.1]octyloxy, azabicylo[3.3.1]nonyloxy,azabicylo[4.3.0]nonyloxy, oxabicylo[2.2.1]heptyloxy,1,1-dioxidotetrahydro-2H-thiopyranyloxy, and1,5,9-triazacyclododecyloxy.

The term “aryl” refers to a monocyclic or fused bicyclic groups having 6to 14 carbon atoms and having at least one aromatic ring that complieswith Hiickel's Rule. Examples of aryl radicals include, but are notlimited to, phenyl, naphthyl, indenyl, azulenyl, fluorenyl, anthracenyl,phenanthrenyl, tetrahydronaphthyl, indanyl, phenanthridinyl and thelike. Unless otherwise indicated, the term “aryl” also includes eachpossible positional isomer of an aromatic hydrocarbon radical, such asin 1-naphthyl, 2-naphthyl, 5-tetrahydronaphthyl, 6-tetrahydronaphthyl,1-phenanthridinyl, 2-phenanthridinyl, 3-phenanthridinyl,4-phenanthridinyl, 7-phenanthridinyl, 8-phenanthridinyl,9-phenanthridinyl and 10-phenanthridinyl.

As used herein, the term “heteroaryl” refers to an aromatic ring systemcontaining carbon(s) and at least one heteroatom selected from nitrogen,oxygen and sulfur, including

N-oxides. Heteroaryl may be monocyclic or polycyclic, substituted orunsubstituted. A monocyclic heteroaryl group may have 1 to 4 heteroatomsin the ring, while a polycyclic heteroaryl may contain 1 to 8heteroatoms. Bicyclic heteroaryl rings may contain from 8 to 10 memberatoms. Monocyclic heteroaryl rings may contain from 5 to 6 member atoms(carbons and heteroatoms). Exemplary 5- to 6-membered heteroarylsinclude, but are not limited to, furanyl, thiophenyl, thienyl, pyrrolyl,imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, oxazolyl, isoxazolyl,1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, thiadiazolyl, isothiazolyl,tetrazolyl, pyridinyl, pyridazinyl, pyrazinyl, pyrimidinyl, andtriazinyl. Other exemplary heteroaryl groups include, but are notlimited to benzofuranyl, isobenzofuryl, 2,3-dihydrobenzofuryl,1,3-benzodioxolyl, dihydrobenzodioxinyl, benzothienyl, indolizinyl,indolyl, isoindolyl, indolinyl, isoindolinyl, benzimidazolyl,dihydrobenzimidazolyl, benzoxazolyl, dihydrobenzoxazolyl, benzthiazolyl,benzoisothiazolyl, dihydrobenzoisothiazolyl, indazolyl,pyrrolopyridinyl, pyrrolopyrimidinyl, imidazopyridinyl,imidazopyrimidinyl, pyrazolopyridinyl, pyrazolopyrimidinyl,benzoxadiazolyl, benzthiadiazolyl, benzotriazolyl, triazolopyridinyl,purinyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl,tetrahydroisoquinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl,quinazolinyl, 1,5-naphthyridinyl, 1,6-naphthyridinyl,1,7-naphthyridinyl, 1,8-naphthyridinyl, and pteridinyl.

As used herein, the term “cyano” refers to the group —CN.

As used herein, the term “optionally” means that the subsequentlydescribed event(s) may or may not occur, and includes both event(s) thatoccur and event(s) that do not occur.

As used herein, unless otherwise defined, the phrase “optionallysubstituted” or variations thereof denote an optional substitution,including multiple degrees of substitution, with one or more substituentgroup. The phrase should not be interpreted as duplicative of thesubstitutions herein described and depicted.

As used herein, the term “treatment” refers to alleviating the specifiedcondition, eliminating or reducing one or more symptoms of thecondition, slowing or eliminating the progression of the condition, andpreventing or delaying the reoccurrence of the condition in a previouslyafflicted or diagnosed patient or subject.

As used herein, the term “effective amount” means that amount of a drugor pharmaceutical agent that will elicit the biological or medicalresponse of a tissue, system, animal, or human that is being sought, forinstance, by a researcher or clinician.

The term “therapeutically effective amount” means any amount which, ascompared to a corresponding subject who has not received such amount,results in improved treatment, healing, prevention, or amelioration of adisease, disorder, or side effect, or a decrease in the rate ofadvancement of a disease or disorder. The term also includes within itsscope amounts effective to enhance normal physiological function. Foruse in therapy, therapeutically effective amounts of a compound ofFormula (I), as well as salts thereof, may be administered as the rawchemical. Additionally, the active ingredient may be presented as apharmaceutical composition.

Compound Preparation Abbreviations

-   Boc₂O di-tent-butyl dicarbonate-   CaCl₂ calcium (II) chloride-   CH₃C1 chloroform-   CH₃CN acetonitrile-   DCE 1,2-dichloroethane-   DCM dichloromethane-   DMAP 4-dimethylaminopyridine-   DMF N,N-dimethylformamide-   DMSO dimethylsulfoxide-   EtOAc ethyl acetate-   EDC N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride-   ES electrospray-   Et₂O diethyl ether-   EtOAc ethyl acetate-   EtOH ethanol-   h hour(s)-   HCl hydrochloric acid-   H₂O water-   HOAt 1-hydroxy-7-azabenzotriazole-   HOBt 1-hydroxybenzotriazole-   HPLC high-performance liquid chromatography-   LCMS liquid chromatography mass spectrometry-   MeOH methanol-   MgCl₂ magnesium chloride-   MgSO₄ magnesium sulfate-   min minute(s)-   MS mass spectrometry-   NaBH₄ sodium borohydride-   NaBH(OAc)₃ sodium triacetoxyborohydride-   Na₂CO₃ sodium carbonate-   NaHCO₃ sodium bicarbonate-   NaOH sodium hydroxide-   Na₂S₂O₃ sodium thiosulfate-   NBS N-bromosuccinimide-   NH₃ ammonia-   NH₄C1 ammonium chloride-   NH₄OH ammonium hydroxide-   NIS N-iodosuccinimide-   NMM N-methylmorpholine-   PyBroP® bromotripyrrolidinophosphonium hexafluorophosphate-   sat. saturated-   TBME tent-butyl methyl ether-   THF tetrahydrofuran-   TFA trifluoroacetic acid

Generic Synthesis Schemes

The compounds of this invention may be made by a variety of methods,including well-known standard synthetic methods. Illustrative generalsynthetic methods are set out below and then specific compounds of theinvention are prepared in the working examples. The skilled artisan willappreciate that if a substituent described herein is not compatible withthe synthetic methods described herein, the substituent may be protectedwith a suitable protecting group that is stable to the reactionconditions. The protecting group may be removed at a suitable point inthe reaction sequence to provide a desired intermediate or targetcompound. In all of the schemes described below, protecting groups forsensitive or reactive groups are employed where necessary in accordancewith general principles of synthetic chemistry. Protecting groups aremanipulated according to standard methods of organic synthesis (T. W.Green and P. G. M. Wuts, (1991) Protecting Groups in Organic Synthesis,John Wiley & Sons, incorporated by reference with regard to protectinggroups). These groups are removed at a convenient stage of the compoundsynthesis using methods that are readily apparent to those skilled inthe art. The selection of processes as well as the reaction conditionsand order of their execution shall be consistent with the preparation ofcompounds of the present invention. Starting materials are commerciallyavailable or are made from commercially available starting materialsusing methods known to those skilled in the art.

The compounds of Formula (I) can be prepared according to Scheme 1 oranalogous methods. An appropriately substituted 3-aminoacrylonitrile iscondensed with an appropriately substituted 1,3-dioxin-4-one withheating to produce a 1,4-dihydropyridin-4-one. Reduction of the nitrileunder the appropriate conditions, such as with a Raney nickel catalystin a hydrogen atmosphere, followed by coupling of the resultant aminewith an appropriately substituted carboxylic acid affords compounds ofFormula (I).

The compounds of Formula (I) can also be prepared according to Scheme 2or analogous methods. Successive reductive alkylations of anappropriately substituted 5-amino-thiophene-3-carboxylate (or itsregioisomer) with appropriately substituted aldehydes or ketonesfurnishes the substituted amines. Saponification of the ester, followedby coupling of the resultant carboxylic acid with an appropriatelysubstituted amine affords compounds of Formula (I).

Experimentals

The following guidelines apply to all experimental procedures describedherein. All reactions were conducted under a positive pressure ofnitrogen using oven-dried glassware, unless otherwise indicated.Temperatures designated are external (i.e. bath temperatures), and areapproximate. Air and moisture-sensitive liquids were transferred viasyringe. Reagents were used as received. Solvents utilized were thoselisted as “anhydrous” by vendors. Molarities listed for reagents insolutions are approximate, and were used without prior titration againsta corresponding standard. All reactions were agitated by stir bar,unless otherwise indicated. Heating was conducted using heating bathscontaining silicon oil, unless otherwise indicated. Reactions conductedby microwave irradiation (0-400 W at 2.45 GHz) were done so using aBiotage Initiator™ 2.0 instrument with Biotage microwave EXP vials(0.2-20 mL) and septa and caps. Irradiation levels utilized (i.e. high,normal, low) based on solvent and ionic charge were based on vendorspecifications. Cooling to temperatures below −70° C. was conductedusing dry ice/acetone or dry ice/2-propanol. Magnesium sulfate andsodium sulfate used as drying agents were of anhydrous grade, and wereused interchangeably. Solvents described as being removed “in vacuo” or“under reduced pressure” were done so by rotary evaporation.

Preparative normal phase silica gel chromatography was carried out usingeither a Teledyne ISCO CombiFlash Companion instrument with RediSep orISCO Gold silica gel cartridges (4 g-330 g), or an Analogix IF280instrument with SF25 silica gel cartridges (4 g-3-00 g), or a BiotageSP1 instrument with HP silica gel cartridges (10 g-100 g). Purificationby reverse phase HPLC was conducted using a YMC-pack column (ODS-A 75×30mm) as solid phase, unless otherwise noted. A mobile phase of 25 mL/minA (CH₃CN-0.1% TFA): B (water-0.1% TFA), 10-80% gradient A (10 min) wasutilized with UV detection at 214 nM, unless otherwise noted.

A PE Sciex API 150 single quadrupoR^(c) mass spectrometer (PE Sciex,Thornhill, Ontario, Canada) was operated using electrospray ionizationin the positive ion detection mode. The nebulizing gas was generatedfrom a zero air generator (Balston Inc., Haverhill, Mass., USA) anddelivered at 65 psi and the curtain gas was high purity nitrogendelivered from a Dewar liquid nitrogen vessel at 50 psi. The voltageapplied to the electrospray needle was 4.8 kV. The orifice was set at 25V and mass spectrometer was scanned at a rate of 0.5 scan/sec using astep mass of 0.2 amu and collecting profile data.

Method A LCMS. Samples were introduced into the mass spectrometer usinga CTC PAL autosampler (LEAP Technologies, Carrboro, NC) equipped with ahamilton 10 uL syringe which performed the injection into a Valco10-port injection valve. The HPLC pump was a Shimadzu LC-10ADvp(Shimadzu Scientific Instruments, Columbia, Md.) operated at 0.3 mL/minand a linear gradient 4.5% A to 90% B in 3.2 min. with a 0.4 min. hold.The mobile phase was composed of 100% (H₂O 0.02% TFA) in vessel A and100% (CH₃CN 0.018% TFA) in vessel B. The stationary phase is Aquasil(C18) and the column dimensions were 1 mm×40 mm. Detection was by UV at214 nm, evaporative light-scattering (ELSD) and MS.

Method B, LCMS. Alternatively, an Agilent 1100 analytical HPLC systemwith an LC/MS was used and operated at 1 mL/min and a linear gradient 5%A to 100% B in 2.2 min with a 0.4 min hold. The mobile phase wascomposed of 100% (H₂O 0.02% TFA) in vessel A and 100% (CH₃CN 0.018% TFA)in vessel B. The stationary phase was Zobax (C8) with a 3.5 um particalsize and the column dimensions were 2.1 mm×50 mm. Detection was by UV at214 nm, evaporative light-scattering (ELSD) and MS.

Method C, LCMS. Alternatively, an MDSSCIEX API 2000 equipped with acapillary column of (50×4.6 mm, 5 μm) was used. HPLC was done onAgilent-1200 series UPLC system equipped with column Zorbax SB-C18(50×4.6 mm, 1.8 μm) eluting with CH₃CN: ammonium acetate buffer. Thereactions were performed in the microwave (CEM, Discover).

¹H-NMR spectra were recorded at 400 MHz using a Bruker AVANCE 400 MHzinstrument, with ACD Spect manager v. 10 used for reprocessing.Multiplicities indicated are: s=singlet, d=doublet, t=triplet,q=quartet, quint=quintet, sxt=sextet, m=multiplet, dd=doublet ofdoublets, dt=doublet of triplets etc. and br indicates a broad signal.All NMRs in DMSO-d₆ unless otherwise noted.

Analytical HPLC: Products were analyzed by Agilent 1100 AnalyticalChromatography system, with 4.5×75 mm Zorbax XDB-C18 column (3.5 um) at2 mL/min with a 4 min gradient from 5% CH₃CN (0.1% formic acid) to 95%CH₃CN (0.1% formic acid) in H₂O (0.1% formic acid) and a 1 min hold.

Intermediates Intermediate 1

3-(Aminomethyl)-2,6-dimethylpyridin-4(1H)-one hydrochloride

a) 2,6-Dimethyl-4-oxo-1,4-dihydropyridine-3-carbonitrile

A 250 mL round bottom flask was charged with 3-aminobut-2-enenitrile(10.00 g, 122 mmol), 2,2,6-trimethyl-4H-1,3-dioxin-4-one (32.4 mL, 244mmol), and a magnetic stir bar. The flask was equipped with a refluxcondenser and a CaC1₂ tube and the reaction mixture was heated at 130°C. for 1 h. The reaction was allowed to cool room temperature and wasdiluted with EtOAc (100 mL). The solid that formed was collected, washedwith EtOAc (20 mL), and dried to give2,6-dimethyl-4-oxo-1,4-dihydropyridine-3-carbonitrile (3.5 g, 19.4%) asa beige soild. ¹H-NMR (400 MHz, DMSO-d₆) δ11.94 (br. s., 1H), 6.04 (s,1H), 2.41 (s, 3H), 2.21 (s, 3H). MS(ES) [M+H]⁺ 148.9.

b) 3-(Aminomethyl)-2,6-dimethylpyridin-4(1H)-one hydrochloride

A 250 mL Erlenmeyer flask was charged with2,6-dimethyl-4-oxo-1,4-dihydropyridine-3-carbonitrile (360 mg, 2.430mmol) and cold 2 M NH₃ in EtOH (39.5 mL, 79 mmol). EtOH (40 mL) wasadded to solubilize the remaining reactant. The solution was passedthrough a Raney Ni cartridge on a continuous flow hydrogenation reactor(40 psi, 40° C., 1 mL/min) for 16 h. The reaction solvent was removed invacuo and the residue was dissolved in EtOH (1 mL) and CHCl₃ (15 mL),then concentrated in vacuo. The residue was dissolved and concentratedin CHCl₃ (2×15 mL) and DCM (15 mL). The sticky residue was suspended inEt₂O (30 mL) and treated with 4 M HC1 in 1,4-dioxanes (10.63 mL, 42.5mmol). The suspension was stirred at room temperature overnight, afterwhich time the white solid was collected via vacuum filtration, washedwith Et₂O (10 mL), and dried under high vacuum to give3-(aminomethyl)-2,6-dimethylpyridin-4(11/)-one hydrochloride (200 mg,80%) as a white solid. ¹H NMR (400 MHz, CDC1₃) 6 6.26 (br. s., 1H), 3.90(br. s., 2H), 2.64 (s, 1H), 2.38 (s, 3H), 2.32 (s, 3H), 1.22-1.30 (m,2H). MS(ES) [M+H]⁺ 152.9.

Intermediate 2

3-Amino-5-(aminomethyl)-2,6-dimethylpyridin-4(1H)-one

a) 2,6-Dimethyl-3-nitropyridin-4(1H)-one

To a cooled (0° C.) solution of sulfuric acid (4.0 mL, 16.24 mmol) wasadded nitric acid (fuming) (4.0 mL, 16.24 mmol) slowly over 5 min. Themixture was stirred for 30 min, at which time was added2,6-dimethylpyridin-4(1H)-one (2.0 g, 16.24 mmol) in portions over 5min. The reaction was allowed to warm to ambient temperature and thenheated at 100° C. for 2 h, at which time it was allowed to cool back toambient temperature. The reaction was poured onto ice (33 g) and themixture was cooled in an ice bath. 8 M NaOH was added and the pHadjusted to 5.3. The solids were filtered and collected to provide2,6-dimethyl-3-nitropyridin-4(1H)-one (440 mg, 15.79%). ¹H NMR (DMSO-d6)δ 11.84 (br. s., 1H), 6.18 (s, 1H), 2.28 (s, 3H), 2.22 (s, 3H). MS(ES)[M+H]⁺ 168.99.

b) 3-Iodo-2,6-dimethyl-5-nitropyridin-4(1H)-one

To a mixture 2,6-dimethyl-3-nitropyridin-4(1H)-one (0.44 g, 2.62 mmol)in acetic acid (13 mL) was NIS (0.765 g, 3.40 mmol). The reaction washeated at 105° C. for 2 h. The reaction was then quenched with ice/water(100 mL) and 0.1 M Na₂S₂O₃ (2 mL) was added. The solids were filteredand collected to provide 3-iodo-2,6-dimethyl-5-nitropyridin-4(11/)-one(596 mg, 69.7%). ¹H NMR (DMSO-d6) δ 12.32 (br. s., 1H), 2.28-2.31 (m,3H). MS(ES) [M+H]⁺ 294.9.

c) 2,6-Dimethyl-5-nitro-4-oxo-1,4-dihydropyridine-3-carbonitrile

A mixture of 3-iodo-2,6-dimethyl-5-nitropyridin-4(1H)-one (0.59 g, 2.006mmol) and copper(I) cyanide (0.359 g, 4.01 mmol) inN-methyl-2-pyrrolidone (10 mL) was heated at 125° C. for 2 h. Thereaction was then quenched with ice/sat. NH₄Cl (100 mL) and cooled in anice bath for 1 h. The solids were filtered and collected to provide2,6-dimethyl-5-nitro-4-oxo-1,4-dihydropyridine-3-carbonitrile (176 mg,40.9%). ¹H NMR (DMSO-d6) δ 12.78 (br. s., 1H), 2.44-2.48 (m, 3H),2.30-2.38 (m, 3H). MS(ES) [M+H]⁺ 193.9.

d) 1,1-Dimethylethyl{[5-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2,6-dimethyl-4-oxo-1,4-dihydro-3-pyridinyl]methyl}carbamate

To a cooled (0° C.) solution of2,6-dimethyl-5-nitro-4-oxo-1,4-dihydropyridine-3-carbonitrile (1.6 g,6.21 mmol) in MeOH (60 mL) were added Boc₂O (4.33 mL, 18.64 mmol) andnickel chloride-hexahydrate (0.738 g, 3.11 mmol), followed by NaBH₄(2.350 g, 62.1 mmol) slowly over about 2 min. The reaction was stirredfor 10 min at 0° C., then was stirred at ambient temperature overnight.To the mixture was added diethylenetriamine (2.70 mL, 24.85 mmol). Thereaction was stirrred for 1 h, at which time it was quenched with sat.NaHCO₃, extracted with 4:1 EtOAc/THF, and concentrated. The residue wasdissolved in MeOH/DCM, adsorbed onto silica gel, and purified by columnchromatography (12 g silica column; Gradient of B: 10-85%, A: DCM, B:10% MeOH in DCM) to provide 1,1-dimethylethyl {[5-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2,6-dimethyl-4-oxo-1,4-dihydro-3-pyridinyl]methyl}carbamate(880 mg, 38.6%). ¹H NMR (DMSO-d6) δ 11.08 (s, 1H), 7.57 (br. s., 1H),6.45 (t, J=5.1 Hz, 1H), 3.94 (d, J=5.3 Hz, 2H), 2.24-2.33 (m, 2H), 2.08-2.16 (m, 4H), 1.40 (s, 12H), 1.33-1.37 (m, 7H). MS(ES) [M+H]⁺ 368.2.

e) 3-Amino-5-(aminomethyl)-2,6-dimethylpyridin-4(1H)-onebis-hydrochloride

To a solution of 4 M HCl/dioxane (10.18 mL, 40.7 mmol) was added slowlya solution of 1,1-dimethylethyl{[5-({[(1,1-dimethylethyl)oxy]carbonyl}amino)-2,6-dimethyl-4-oxo-1,4-dihydro-3-pyridinyl]methyl}carbamate(880 mg, 2.395 mmol) in EtOAc (25 mL) and CH₃Cl (5 mL). The reaction washeated at 40° C. for 4 h, at which time it was diluted with Et₂O (100mL) and concentrated to provide3-amino-5-(aminomethyl)-2,6-dimethylpyridin-4(1H)-one (686 mg, 119%) asits bis-hydrochloride salt. ¹H NMR (DMSO-d6) δ 8.17 (br. s., 3H),3.82-3.92 (m, 2H), 2.52 (s, 3H), 2.44 -2.48 (m, 3H), 2.35-2.40 (m, 2H).MS(ES) [M+H]⁺ 168.0.

EXAMPLES Example 1

N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxamide

a) Methyl5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxylate

To a solution of commercially available methyl5-amino-4-methylthiophene-3-carboxylate (500 mg, 2.92 mmol) in DCE (10mL) were added acetic acid (0.334 mL, 5.84 mmol) anddihydro-2H-pyran-4(3H)-one (322 mg, 3.21 mmol). The reaction wasmaintained at room temperature for 20 min, at which time NaBH(OAc)₃(2476 mg, 11.68 mmol) was added portion wise. The reaction was stirredfor 90 min. LCMS showed the reaction was complete. To the reactionmixture was added acetaldehyde (0.198 mL, 3.50 mmol) and NaBH(OAc)₃(˜500 mg). The reaction was stirred for 1 h, at which time it wasquenched with sat. NaHCO₃, extracted with DCM, and concentrated. Theresidue was purified by silica gel chromatography (Varian 971IF, 0-50%EtOAc/hexanes, SF25-40g, 15 minutes) to give methyl5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxylate(750 mg, 89%) as a tan oil. ¹H NMR (400 MHz, CDCl3) δ 7.95 (s, 1 H) 3.99(dt, J=10.36, 2.02 Hz, 2 H) 3.86 (s, 3 H) 3.37 (td, J=11.87, 2.02 Hz,2H) 2.94-3.04 (m, 3 H) 2.35 (s, 3 H) 1.75-1.82 (m, 2 H) 1.58-1.62 (m, 2H) 0.96 (t, J=7.20 Hz, 3 H). MS(ES) [M+H]⁺ 284.0.

b)N-((2,6-Dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxamide

To a solution of methyl5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxylate(150 mg, 0.529 mmol) in MeOH was added 5 N NaOH (0.529 mL, 2.65 mmol).The reaction was stirred at 40° C. for 2 h. The reaction was allowed tocool to room temperature and was diluted with 6 N HCl (0.441 mL, 2.65mmol). The mixture was concentrated, diluted with DCM, and concentratedagain.

To a solution of the crude residue in DMSO (5 mL) was added NMM (0.291mL, 2.65 mmol), 3-(aminomethyl)-2,6-dimethylpyridin-4(1H)-one (97 mg,0.635 mmol), EDC (122 mg, 0.635 mmol), HOBt (97 mg, 0.635 mmol), andmore NMM (0.291 mL, 2.65 mmol). The reaction was maintained at roomtemperature for 75 h, at which time it was purified by reverse phaseHPLC (Gilson; 5-60% CH₃CN/water +0.1% TFA, YMC ODS-A C18 column 75×30 mmID S-5 um). The residue was partitioned between EtOAc/MeOH and sat.aqueous NaHCO₃. The organic layer was concentrated, diluted with DCM,and concentrated again to provideN-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxamide(95 mg, 42.3%) as an off-white foam solid. ¹H NMR (400 MHz, DMSO-d₆) δ11.02 (br. s., 1 H) 7.99 (br. s., 1 H) 7.64 (s, 1 H) 5.88 (br. s., 1 H)4.18 (d, J=5.31 Hz, 2 H) 3.83 (d, J=14.15 Hz, 2H) 3.21-3.29 (m, 2 H)2.89-2.95 (m, 3 H) 2.32 (s, 3 H) 2.14 -2.17 (m, 6 H) 1.68 (dd, J=12.25,1.89 Hz, 2 H) 1.33-1.44 (m, 2 H) 0.85-0.89 (m, 3 H). MS(ES) [M+H]⁺404.1.

Example 2

N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide

a) Methyl 5 -(((cis)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthi ophene-3-carboxylate andMethyl 5-(((trans)-4-(dimethyl amino)cy clohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate

To a solution of methyl 5-amino-4-methylthiophene-3-carboxylate (800 mg,4.67 mmol) in DCE (25 mL) were added acetic acid (0.535 mL, 9.34 mmol)and 4-(dimethylamino)cyclohexanone (660 mg, 4.67 mmol). The reaction wasmaintained at room temperature for 40 min, at which time NaBH(OAc)₃(3961 mg, 18.69 mmol) was added portion-wise. The mixture was stirred atroom temperature for 30 min, at which time acetaldehyde (0.317 mL, 5.61mmol) and more NaBH(OAc)₃ (990 mg, 4.67 mmol) were added. The reactionwas stirred for 1 h, at which time it was quenched with NaHCO₃,extracted with DCM, and concentrated. The crude residue was purified byreverse phase Gilson HPLC (10-70% CH₃CN/water +0.1% TFA, YMC ODS-A C18Column 75×30 mm ID S-5 um, 12 nM Column 6 min). The semi pure residuewas partitioned between EtOAc and NaHCO₃ (sat. aq). The organics wereconcentrated, diluted with DCM and evaporated to provide methyl5-((4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(mixture of cis and trans isomers) as a brown oil. MS(ES) [M+H]⁺ 325.1.

The cis and trans isomers were separated by chiral HPLC (Gilson; IC20×250 mm column; 20 mL/min; 5:95 EtOH:Heptane) to give:

Methyl5-(((cis)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(180 mg, 11.87%). ¹HNMR (400 MHz, CDCl₃) δ 7.93 (s, 1 H) 3.85 (s, 3 H)2.94-3.00 (m, 3 H) 2.37 (s, 3 H) 2.28 (s, 6 H) 2.16 (br. s., 1 H)1.83-1.91 (m, 2 H) 1.71-1.80 (m, 2 H) 1.44-1.51 (m, 4 H) 0.93 (t, J=7.20Hz, 3 H). MS(ES) [M+H]⁺ 325.2.

Methyl 5 -(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(387 mg, 25.5%). ¹H NMR (400 MHz, CDC1₃) δ 7.91 (s, 1 H) 3.85 (s, 3 H)2.98 (q, J=7.07 Hz, 2 H) 2.31 (s, 3 H) 2.24-2.28 (m, 6H) 2.10 (m,J=10.89, 7.36, 3.51, 3.51 Hz, 1 H) 1.98 (d, J=10.11 Hz, 2H) 1.89-1.93(m, 2 H) 1.20 -1.32 (m, 4 H) 0.93 (t, J=7.07 Hz, 3 H). MS(ES) [M+H]⁺325.2.

b)5-(((trans)-4-(Dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylicacid

To a solution of methyl5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(380 mg, 1.71 mmol) in MeOH was added 5 N NaOH (1.71 mL, 5.86 mmol). Themixture was stirred overnight at 37° C., at which time 6 N HCl (1.05 mL)was added. The mixture was concentrated to provide5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylicacid (690 mg, 95%) as a solid. MS(ES) [M+H]⁺ 311.2.

c)N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide

To a solution of5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylicacid (150 mg, 0.483 mmol) in DMSO (5 mL) were added NMM (0.266 mL, 2.416mmol) and 3-(aminomethyl)-2,6-dimethylpyridin-4(1H)-one (88 mg, 0.580mmol), followed by EDC (185 mg, 0.966 mmol), HOBt (148 mg, 0.966 mmol),and more NMM (0.266 mL, 2.416 mmol). The reaction was stirred at roomtemperature for 12 h. The reaction was purified directly by reversephase HPLC (Gilson, 5-55% CH₃CN/water+0.1% TFA, YMC ODS-A C18 column75×30mm ID S-5 um, 12 nM column). The residue was partitioned betweenEtOAc/MeOH and 0.1N NaOH. The organics were concentrated, diluted withDCM, and concentrated again to provideN-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide(80 mg, 35.4%) as an off white foam-solid. ¹H NMR (600 MHz, DMSO-d6) δ0.85 (t, J=6.80 Hz, 3 H) 1.11-1.24 (m, 4 H) 1.77 (d, J=9.82 Hz, 2 H)1.86 (d, J=9.82 Hz, 2 H) 2.05 (d, J=8.69 Hz, 1 H) 2.12 (br. s., 3 H)2.14 (br. s., 6 H) 2.15 (br. s., 3 H) 2.31 (s, 3 H) 2.66 (br. s., 1 H)2.91 (q, J=6.80 Hz, 2 H) 4.17 (d, J=4.15 Hz, 2 H) 5.87 (br. s., 1 H)7.59 (s, 1 H) 7.94 (br. s., 1 H) 11.01 (br. s., 1 H). MS(ES) [M+H]⁺445.3. Additional NMR studies (NOE) confirmed the trans geometry.

Example 3

N-((5-amino-2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide

To a solution of5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylicacid (170 mg, 0.274 mmol) in DMSO (3 mL) were added3-amino-5-(aminomethyl)-2,6-dimethylpyridin-4(1H)-one bis-hydrochloride(79 mg, 0.324 mmol), HOAt (45 mg, 0.329 mmol), and EDC (63 mg, 0.329mmol), followed by NMIVI (2.196 mL, 19.97 mmol). The reaction wasstirred at ambient temperature for 2 h, at which time water (2 mL) wasadded and the reaction was allowed to stand overnight. The reaction waspurified directly by prep HPLC (Gilson; Gradient of B: 8-45%, A:water+0.1% TFA, B: CH₃CN+0.1% TFA). The resulting residue was dissolvedin MeOH/DCM and TEA (0.3 ml), adsorbed onto silica gel, and purified bycolumn chromatography (4 g silica column; Gradient of B: 25-100%, A:DCM, B: 90/10/1 of CHCl₃/MeOH/NH₄OH). The residue solidified fromevaporation with CH₃CN, followed by TBME to provideN45-amino-2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide(51 mg, 39%). ¹H NMR (DMSO-d6) δ 10.82 (s, 1H), 8.01 (t, J=5.1 Hz, 1H),7.58 (s, 1H), 4.15-4.30 (m, 4H), 2.90 (q, J=6.9 Hz, 2H), 2.65 (d, J=3.3Hz, 1H), 2.29 (s, 3H), 2.07-2.18 (m, 12H), 2.01 (br. s., 1H), 1.85 (d,J=10.1 Hz, 2H), 1.76 (d, J=9.9 Hz, 2H), 1.12-1.25 (m, 4H), 0.84 (t,J=6.9 Hz, 3H). MS(ES) [M+H]⁺ 460.3.

Example 4

2-Bromo-N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide

a) Methyl2-bromo-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate

A solution of methyl5-((4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(5.21 g, 16.06 mmol) in DMF (32 mL) was treated with NBS (4.00 g, 22.48mmol) and stirred under argon at room temperature for 30 min. Thereaction was diluted with water (10 mL) and saturated Na₂CO₃ solution(pH 9, 1 mL). The resulting suspension was extracted with EtOAc (3×75mL). The organics were washed with water (50 mL), dried over MgSO₄ andconcentrated onto silica. The material was purified on an ISCO®Combiflash Rf on 24 g silica using a gradient of CHCl₃:MeOH w/1% NH₄OH(0-15%). The product fractions were combined and concentrated to affordmethyl2-bromo-5-((4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(3.83 g) as a mixture of cis- and trans-cyclohexyl isomers. The mixturewas then separated by HPLC (Chiralpak AD-H, 5 microns, 50 mm×250 mm,97:3:0.1 CH₃CN:isopropylalcohol: isopropylamine, 100 mL/min) to affordmethyl2-bromo-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(2.64 g, 6.41 mmol, 39.9% yield) as an orange tar. ¹H NMR (400 MHz,CDC1₃) 6 3.90 (s, 3 H) 2.94 (q, J=7.07 Hz, 2 H) 2.64-2.77 (m, 1 H) 2.27(s, 6 H) 2.21 (s, 3 H) 2.05-2.16 (m, 1 H) 1.89-2.00 (m, 4 H) 1.16-1.34(m, 4 H) 0.96 (t, J=7.07 Hz, 3 H). MS(ES) [M+H]⁺ 403, 405.

b)2-Bromo-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylicacid

A solution of methyl2-bromo-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylate(0.150 g, 0.372 mmol) in MeOH (1.5 mL) and THF (0.300 mL) was treatedwith 6 M NaOH (0.310 mL, 1.859 mmol). The reaction was heated in a 45°C. oil bath for 18 h. The reaction was neutralized with 5 M HCl (0.372mL, 1.859 mmol) and concentrated to dryness. MS(ES) [M+H]⁺ 389.

c) 2-Bromo—N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide

A mixture of2-bromo-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxylicacid (0.145 g, 0.372 mmol),3-(aminomethyl)-2,6-dimethylpyridin-4(1H)-one hydrochloride (0.070 g,0.372 mmol), and DMAP (4.55 mg, 0.037 mmol) in DMSO (2 mL) was addedN,N-diisopropylethylamine (0.260 mL, 1.490 mmol), followed by PyBroP®(0.191 g, 0.410 mmol). The reaction was stirred overnight under argon atroom temperature. LCMS showed a 2:1 mixture of product to startingmaterial. The reaction was diluted with MeOH (1 mL) and filtered.Purification of the residue by preparative HPLC (Waters Sunfire 30×150mm column; 10-40% CH₃CN/0.1% TFA in water) gave2-bromo-N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-54((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide(54 mg, 26%). ¹H NMR (400 MHz, DMSO-d₆) δ 11.03 (s, 1 H) 8.20 (t, J=4.89Hz, 1 H) 5.87 (s, 1 H) 4.20 (d, J=4.77 Hz, 2 H) 2.88 (q, J=6.86 Hz, 2 H)2.62 -2.71 (m, 1 H) 2.31 (s, 3 H) 2.16 (s, 3 H) 2.12 (s, 6 H) 1.98-2.07(m, 1 H) 1.92 (s, 3 H) 1.85 (d, J=10.04 Hz, 2 H) 1.77 (d, J=11.04 Hz, 2H) 1.07-1.26 (m, 4 H) 0.89 (t, J=7.03 Hz, 3 H). MS(ES) [M+H]⁺ 523.2.

Assay Protocol 1

Compounds contained herein were evaluated for their ability to inhibitthe methyltransferase activity of EZH2 within the PRC2 complex. HumanPRC2 complex was prepared by co-expressing each of the 5 member proteins(FLAG-EZH2, EED, SUZ12, RbAp48, AEBP2) in Sf9 cells followed byco-purification. Enzyme activity was measured in a scintillationproximity assay (SPA) where a tritiated methyl group is transferred from3H-SAM to a lysine residue on Histone H3 of a mononucleosome, purifiedfrom HeLa cells. Mononucleosomes were captured on SPA beads and theresulting signal is read on a ViewLux plate reader.

Part A. Compound Preparation

1. Prepare 10 mM stock of compounds from solid in 100% DMSO.

2. Set up an 11-point serial dilution (1:3 dilution, top concentration10 mM) in 100% DMSO for each test compound in a 384 well plate leavingcolumns 6 and 18 for DMSO controls.

3. Dispense 100 nL of compound from the dilution plate into reactionplates (Grenier Bio-One, 384-well, Cat# 784075).

Part B. Reagent Preparation

Prepare the following solutions:

1. 50 mM Tris-HCl, pH 8: Per 1 L of base buffer, combine 1 M Tris-HC1,pH 8 (50 mL) and distilled water (950 mL).

2. 1× Assay Buffer: Per 10 mL of 1× Assay Buffer, combine 50 mMTris-HCl, pH 8 (9958 uL), 1 M MgCl₂ (20 uL), 2 M DTT (20 uL), and 10%Tween-20 (2 uL) to provide a final concentration of 50 mM Tris-HCl, pH8, 2 mM MgCl₂, 4 mM DTT, 0.002% Tween-20.

3. 2× Enzyme Solution: Per 10 mL of 2× Enzyme Solution, combine 1× AssayBuffer and PRC2 complex to provide a final enzyme concentration of 10nM.

4. SPA Bead Suspension: Per 1 mL of SPA Bead Suspension, combine PS-PEIcoated LEADSeeker beads (40 mg) and H₂O (1 mL) to provide a finalconcentration of 40 mg/mL.

5. 2× Substrate Solution: Per 10 mL of 2× Substrate Solution, combine 1×Assay Buffer (9728.55 uL), 800 ug/mL mononucleosomes (125 uL), 1 mM coldSAM (4 uL), and 7.02 uM 3H-SAM (142.45 uL; 0.55 mCi/mL) to provide afinal concentration of 5 ug/mL nucleosomes, 0.2 uM cold SAM, and 0.05 uM3H-SAM. 6. 2.67× Quench/Bead Mixture: Per 10 mL of 2.67× Quench/BeadMixture, combine ddH₂O (9358 uL), 10 mM cold SAM (267 uL), 40 mg/mL BeadSuspension (375 uL) to provide a final concentration of 100 uM cold SAMand 0.5 mg/mL SPA beads.

Part C. Assay Reaction in 384-well Grenier Bio-One Plates

Compound Addition

1. Dispense 100 nL/well of 100× Compound to test wells (as noted above).

2. Dispense 100 nL/well of 100% DMSO to columns 6 & 18 for high and lowcontrols, respectively.

Assay

1. Dispense 5 uL/well of 1× Assay Buffer to column 18 (low controlreactions).

2. Dispense 5 uL/well of 2× Enzyme Solution to columns 1-17, 19-24.

3. Spin assay plates for 1 min at 500 rpm.

4. Stack the assay plates, covering the top plate.

5. Incubate the compound/DMSO with the enzyme for 30 min at roomtemperature.

6. Dispense 5 uL/well of 2× Substrate Solution to columns 1-24.

7. Spin assay plates for ˜1 min at 500 rpm.

8. Stack the assay plates, covering the top plate.

9. Incubate the assay plates at room temperature for 1 hour.

Quench/Bead Addition

1. Dispense 5 uL/well of the 3× Quench/Bead Mixture to columns 1-24.

2. Seal the top of each assay plate with adhesive TopSeal.

3. Spin assay plates for ˜1 min at 500 rpm.

4. Equilibrate the plates for >20 min.

Read plates

1. Read the assay plates on the Viewlux Plate Reader utilizing the 613nm emission filter with a 300 s read time.

Reagent addition can be done manually or with automated liquid handler.*The final DMSO concentration in this assay is 1%.*The positive control is in column 6; negative control is in column 18.*Final starting concentration of compounds is 100 μM.

Results

Percent inhibition was calculated relative to the DMSO control for eachcompound concentration and the resulting values were fit using standardIC₅₀ fitting parameters within the ABASE data fitting software package.

The compounds of Examples 1-3 were generally tested according to theabove or an analogous assay and were found to be inhibitors of EZH2.Specific biological activities tested according to such assays arelisted in the following table. Repeating the assay run(s) may result insomewhat different IC₅₀ values.

EZH2 IC₅₀ Example (nM) 1 316 2 32 3 32

Assay Protocol 2

Compounds contained herein were evaluated for their ability to inhibitthe methyltransferase activity of EZH2 within the PRC2 complex. HumanPRC2 complex was prepared by co-expressing each of the 5 member proteins(FLAG-EZH2, EED, SUZ12, RbAp48, AEBP2) in Sf9 cells followed byco-purification. Enzyme activity was measured in a scintillationproximity assay (SPA) where a tritiated methyl group is transferred from3H-SAM to a lysine residue on a biotinylated, unmethylated peptidesubstrate derived from histone H3. The peptides were captured onstreptavidin-coated SPA beads and the resulting signal was read on aViewLux plate reader.

Part A. Compound Preparation

4. Prepare 10 mM stock of compounds from solid in 100% DMSO.

5. Set up an 11-point serial dilution (1:4 dilution, top concentration10 mM) in 100% DMSO for each test compound in a 384 well plate leavingcolumns 6 and 18 for DMSO controls.

6. Dispense 10 nL of compound from the dilution plate into reactionplates (Corning, 384-well polystyrene NBS, Cat# 3673).

Part B. Reagent Preparation

Prepare the following solutions:

7. 1× Base Buffer, 50 mM Tris-HCl, pH 8, 2 mM MgC1₂: Per 1 L of basebuffer, combine 1 M Tris-HC1, pH 8 (50 mL), 1 M MgC1₂ (2 mL), anddistilled water (948 mL).

8. 1× Assay Buffer: Per 10 mL of 1× Assay Buffer, combine 1× Base Buffer(9.96 mL), 1 M DTT (40 uL), and 10% Tween-20 (1 uL) to provide a finalconcentration of 50 mM Tris-HC1, pH 8, 2 mM MgC1₂, 4 mM DTT, 0.001%Tween-20.

9. 2× Enzyme Solution: Per 10 mL of 2× Enzyme Solution, combine 1× AssayBuffer (9.99 mL) and 3.24 uM EZH2 5 member complex (6.17 uL) to providea final enzyme concentration of 1 nM.

10. SPA Bead Solution: Per 1 mL of SPA Bead Solution, combineStreptavidin coated SPA beads (PerkinElmer, Cat# RPNQ0261, 40 mg) and 1×Assay Buffer (1 mL) to provide a working concentration of 40 mg/mL.

11. 2× Substrate Solution: Per 10 mL of 2× Substrate Solution, combine40 mg/mL SPA Bead Solution (375 uL), 1 mM biotinylated histone H3K27peptide (200 uL), 12.5 uM 3H-SAM (240 uL; 1 mCi/mL), 1 mM cold SAM (57uL), and 1× Assay Buffer (9.13 mL) to provide a final concentration of0.75 mg/mL SPA Bead Solution, 10 uM biotinylated histone H3K27 peptide,0.15 uM 3H-SAM (˜12 uCi/mL 3H-SAM), and 2.85 uM cold SAM.

12. 2.67× Quench Solution: Per 10 mL of 2.67× Quench Solution, combine1× Assay Buffer (9.73 mL) and 10 mM cold SAM (267 uL) to provide a finalconcentration of 100 uM cold SAM.

Part C. Assay Reaction in 384-well Grenier Bio-One Plates

Compound Addition

3. Stamp 10 nL/well of 1000× Compound to test wells (as noted above).

4. Stamp 10 nL/well of 100% DMSO to columns 6 & 18 (high and lowcontrols, respectively).

Assay

10. Dispense 5 uL/well of 1× Assay Buffer to column 18 (low controlreactions).

11. Dispense 5 uL/well of 2× Substrate Solution to columns 1-24 (note:substrate solution should be mixed to ensure homogeneous bead suspensionbefore dispensing into matrix reservoir).

12. Dispense 5 uL/well of 2× Enzyme Solution to columns 1-17, 19 24.

13. Incubate the reaction for 60 min at room temperature.

Quench

5. Dispense 6 uL/well of the 2.67× Quench Solution to columns 1 24.

6. Seal assay plates and spin for ˜1 min at 500 rpm.

7. Dark adapt plates in the ViewLux instrument for 15-60 min.

Read plates

2. Read the assay plates on the Viewlux Plate Reader utilizing the 613nm emission filter or clear filter (300 s exposure).

Reagent addition can be done manually or with automated liquid handler.

Results

Percent inhibition was calculated relative to the DMSO control for eachcompound concentration and the resulting values were fit using standardIC₅₀ fitting parameters within the ABASE data fitting software package.

The compounds of Examples 2-4 were generally tested according to theabove or an analogous assay and were found to be inhibitors of EZH2.Specific biological activities tested according to such assays arelisted in the following table. Repeating the assay run(s) may result insomewhat different IC₅₀ values.

EZH2 IC₅₀ Example (nM) 2 100 3 16 4 160

1. A compound according to Formula (I):

wherein: A is NH; X is O, N, S, CR⁶, or NR⁷; Y is O, N, S, CR⁶, or NR⁷;Z is CR⁵ or NR⁸; wherein when X is O, S, or NR⁷, Y is N or CR⁶ and Z isCR⁵; when Y is O, S, or NR⁷, X is N or CR⁶ and Z is CR⁵; and when Z isNR⁸, Y is N or CR⁶ and X is N or CR⁶; R¹, R², and R³ are eachindependently selected from the group consisting of hydrogen,(C₁-C4)alkoxy, (C₁-C₈)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl-,halo(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl, hydroxy(C₁-C₄)alkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, R^(a)O(O)CNH(C₁-C₄)alkyl-,(C6-C₁₀)bicycloalkyl, heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-,aryl, aryl(C₁-C₄)alkyl, heteroaryl, heteroaryl(C₁-C₄)alkyl, halogen,cyano, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(a)R^(b),—SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b), nitro, —NR^(a)R^(b),—NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a),—NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b),—NR^(a)NR^(a)C(O)R^(b), —NR^(a)NR^(a)C(O)NR^(a)R^(b),—NR^(a)NR^(a)C(O)OR^(a), —OR^(a), —OC(O)R^(a), and —OC(O)NR^(a)R^(b),wherein each (C3-C8)cycloalkyl, (C₆-C₁₀)bicycloalkyl, heterocycloalkyl,aryl, or heteroaryl is optionally substituted 1, 2, or 3 times,independently, by hydroxyl, halogen, nitro, (C₁-C₄)alkyl, cyano,(C₁-C₄)alkoxy, —NR^(a)R^(b) or —CO₂R^(a); R⁴ is selected from the groupconsisting of hydrogen, (C₁-C₃)alkoxy, (C₁-C₃)alkyl, hydroxyl, halogen,cyano, (C₃-C₆)cycloalkyl, heterocycloalkyl, —NR^(a)R^(b),halo(C₁-C₃)alkyl, and hydroxy(C₁-C₃)alkyl; R⁵ is selected from the groupconsisting of (C₄-C₈)alkyl, (C₂-C₈)alkenyl, (C₃-C₈)alkoxy,(C₄-C₈)cycloalkyl, (C₃-C₈)cycloalkyl(C₁-C₂)alkyl-,(C₃-C₈)cycloalkyloxy-, heterocycloalkyl, heterocycloalkyl(C₁-C₂)alkyl-,heterocycloalkyloxy-, aryl, heteroaryl, and —NR^(a)R^(b), wherein said(C₄-C₈)alkyl, (C₂-C₈)alkenyl, (C₃-C₈)alkoxy, (C₄-C₈)cycloalkyl,(C₃-C₈)cycloalkyl(C₁-C₂)alkyl-, (C₃-C₈)cycloalkyloxy-, heterocycloalkyl,heterocycloalkyl(C₁-C₂)alkyl-, heterocycloalkyloxy-, aryl, or heteroarylis optionally substituted 1, 2, or 3 times, independently, by halogen,—OR^(a), —NR^(a)R^(b), —NHCO₂R^(a), nitro, (C₁-C₃)alkyl,R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano,—CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), heterocycloalkyl, aryl, orheteroaryl, wherein said (C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, orheteroaryl is optionally substituted 1 or 2 times, independently, byhalogen, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, —CO(C₁-C₄)alkyl,—CO₂(C₁-C₄)alkyl, —NRaR^(b), —NHCO₂R^(a), hydroxyl, oxo, (C₁-C₄)alkoxy,or (C₁-C4)alkoxy(C₁-C₄)alkyl-; or any 2 optional substituents on said(C₂-C₈)alkenyl taken together with the carbon atom(s) to which they areattached represent a 5-8 membered ring, optionally containing aheteroatom selected from oxygen, nitrogen, and sulfur, wherein said ringis optionally substituted 1 or 2 times, independently, by (C₁-C₄)alkyl,halo(C₁-C₄)alkyl, —CO(C₁-C₄)alkyl, —CO₂(C₁-C₄)alkyl, —NRaR^(b),—NHCO₂R^(a), hydroxyl, oxo, (C₁-C₄)alkoxy, or(C₁-C₄)alkoxy(C₁-C₄)alkyl-; R⁶ is selected from the group consisting ofhydrogen, halogen, (C₁-C₈)alkyl, (C₁-C₄)alkoxy, —B(OH)₂,(C₃-C₈)cycloalkyl, (C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C₆-C₁₀)bicycloalkyl,heterocycloalkyl, heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl,heteroaryl, heteroaryl(C₁-C₄)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(a)R^(b), —SR^(a), —S(O)R^(a),—SO₂R^(a), —SO₂NR^(a)R^(b), nitro, —NR^(a)R^(b),R^(a)R^(b)N(C₁-C₄)alkyl-, —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b),—NR^(a)NR^(a)R^(b), —NR^(a)NR^(a)C(O)R^(b),—NR^(a)NR^(a)C(O)NR^(a)R^(b), —NR^(a)NR^(a)C(O)OR^(a), —OR^(a),—OC(O)R^(a), and —OC(O)NR^(a)R^(b), wherein each cycloalkyl,bicycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is optionallysubstituted 1, 2, or 3 times, independently, by R^(c)-(C₁-C₆)alkyl-O—,R^(c)-(C₁-C₆)alkyl-S—, R^(c)-(C₁-C₆)alkyl-,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heterocycloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl; R⁷ is selected from thegroup consisting of hydrogen, (C₁-C₈)alkyl, (C₃-C₈)cycloalkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C₆-C ₁₀)bicycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl, heteroaryl,heteroaryl(C₁-C₄)alkyl, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b),—C(O)NRNR^(a)R^(b), —SO₂R^(a), —SO₂NR^(a)R^(b), andR^(a)R^(b)N(C₁-C₄)alkyl-, wherein each cycloalkyl, bicycloalkyl,heterocycloalkyl, aryl, or heteroaryl group is optionally substituted 1,2, or 3 times, independently, by R^(c)-(C₁-C₆)alkyl-O—,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b), nitro,—NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heteroc_(y)cloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl; R⁸ is selected from thegroup consisting of (C₄-C₈)alkyl, (C₄-C₈)cycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₂)alkyl-, aryl, and heteroaryl, wherein said(C₄-C₈)alkyl, (C₄-C₈)cycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₂)alkyl-, aryl, or heteroaryl is optionallysubstituted 1, 2, or 3 times, independently, by halogen, —OR^(a),—NR^(a)R^(b), —NHCO₂R^(a), nitro, (C₁-C₃)alkyl,R^(a)R^(b)N(C₁-C₃)alkyl-, R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano,—CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), aryl, or heteroaryl; eachR^(c) is independently —S(O)R^(a), —SO₂R^(a), —NR^(a)R^(b)—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), or —CO₂R^(a); and R^(a) and R^(b)are each independently hydrogen, (C₁-C₄)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl-, (C₃-C₁₀)cycloalkyl, heterocycloalkyl, aryl,aryl(C₁-C₄)alkyl-, heteroaryl(C₁-C₄)alkyl-, or heteroaryl, wherein anysaid cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group isoptionally substituted 1, 2, or 3 times, independently, by halogen,hydroxyl, (C₁-C₄)alkoxy, amino, —NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂,(C₁-C₄)alkyl, halo(C₁-C₄)alkyl, —CO₂H, —CO₂(C₁-C₄)alkyl, —CONH₂,—CONH(C₁-C₄)alkyl, —CON((C₁-C₄)alkyl)₂, —SO₂(C₁-C₄)alkyl, —SO₂NH₂,—SO₂NH(C₁-C₄)alkyl, or —SO₂N((C₁-C₄)alkyl)₂; or R^(a) and R^(b) takentogether with the nitrogen to which they are attached represent a 5-8membered saturated or unsaturated ring, optionally containing anadditional heteroatom selected from oxygen, nitrogen, and sulfur,wherein said ring is optionally substituted 1, 2, or 3 times,independently, by (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, amino,—NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂, hydroxyl, oxo, (C₁-C₄)alkoxy, or(C₁-C₄)alkoxy(C₁-C₄)alkyl-, wherein said ring is optionally fused to a(C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring; or R^(a)and R^(b) taken together with the nitrogen to which they are attachedrepresent a 6- to 10-membered bridged bicyclic ring system optionallyfused to a (C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroarylring; or a pharmaceutically acceptable salt thereof. 2-5. (canceled) 6.The compound according to claim 1 represented by Formula (III)(a):

wherein: R¹, R², and R³ are each independently selected from the groupconsisting of hydrogen, (C₁-C4)alkoxy, (C₁-C₈)alkyl,(C₁-C₄)alkoxy(C₁-C₄)alkyl-, halo(C₁-C₄)alkyl, (C₃-C₈)cycloalkyl,hydroxy(C ₁-C₄)alkyl, (C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, R^(a)0(O)CNH(C₁-C₄)alkyl-, (C₆-C ₁₀)bicycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl, heteroaryl,heteroaryl(C₁-C₄)alkyl, halogen, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —C(O)NR^(a)NR^(a)R^(b), —SR^(a), —S(O)R^(a),—SO₂R^(a), —SO₂NR^(a)R^(b), nitro, —NR^(a)R^(b), —NR^(a)C(O)R^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b),—NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b), —NR^(a)NR^(a)C(O)R^(b),—NR^(a)NR^(a)C(O)NR^(a)R^(b), —NR^(a)R_(a)C(O)OR^(a), —OR^(a),—OC(O)R^(a), and —OC(O)NR^(a)R^(b), wherein each (C₃-C₈)cycloalkyl,(C₆-C ₁₀)bicycloalkyl, heterocycloalkyl, aryl, or heteroaryl isoptionally substituted 1, 2, or 3 times, independently, by hydroxyl,halogen, nitro, (C₁-C₄)alkyl, cyano, (C₁-C₄)alkoxy, —NR^(a)R^(b) or—CO₂R^(a); R⁴ is selected from the group consisting of hydrogen,(C₁-C₃)alkoxy, (C₁-C₃)alkyl, hydroxyl, halogen, cyano,(C₃-C₆)cycloalkyl, heterocycloalkyl, —NR^(a)R^(b), halo(C₁-C₃)alkyl, andhydroxy(C₁-C₃)alkyl; R⁵ is selected from the group consisting of(C₄-C₈)alkyl, (C₃-C₈)alkoxy, (C₄-C₈)cycloalkyl, (C₃-C₈)cycloalkyloxy-,heterocycloalkyl, heterocycloalkyloxy-, aryl, heteroaryl, and—NR^(a)R^(b), wherein said (C₄-C₈)alkyl, (C₃-C₈)alkoxy,(C₄-C₈)cycloalkyl, (C₃-C₈)cycloalkyloxy-, heterocycloalkyl,heterocycloalkyloxy-, aryl, or heteroaryl is optionally substituted 1,2, or 3 times, independently, by halogen, —OR^(a), —NR^(a)R^(b),—NHCO₂R^(a), nitro, (C₁-C₃)alkyl, R^(a)R^(b)N(C₁-C₃)alkyl-,R^(a)O(C₁-C₃)alkyl-, (C₃-C₈)cycloalkyl, cyano, —CO₂R^(a),—C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), aryl, or heteroaryl; R⁶ is selectedfrom the group consisting of hydrogen, halogen, (C₁-C₈)alkyl,(C₁-C₄)alkoxy, -B(OH)₂, (C₃-C₈)cycloalkyl,(C₃-C₈)cycloalkyl(C₁-C₄)alkyl-, (C₆-C₁₀)bicycloalkyl, heterocycloalkyl,heterocycloalkyl(C₁-C₄)alkyl-, aryl, aryl(C₁-C₄)alkyl, heteroaryl,heteroaryl(C₁-C₄)alkyl, cyano, —C(O)R^(a), —CO₂R^(a), —C(O)NR^(a)R^(b),—C(O)NR^(a)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), R^(a)R^(b)N(C₁-C₄)alkyl-, —NR^(a)C(O)R^(b),—NR^(a)C(O)NR^(a)R^(b), —NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b),—NR^(a)SO₂NR^(a)R^(b), —NR^(a)NR^(a)R^(b), —NR^(a)NR^(a)C(O)R^(b),—NR^(a)NR^(a)C(O)NR^(a)R^(b), —NR^(a)NR^(a)C(O)OR^(a), —OR^(a),—OC(O)R^(a), and —OC(O)NR^(a)R^(b), wherein each cycloalkyl,bicycloalkyl, heterocycloalkyl, aryl, or heteroaryl group is optionallysubstituted 1, 2, or 3 times, independently, by R^(c)-(C₁-C₆)alkyl-,(C₁-C₄)alkyl-heterocycloalkyl-, halogen, (C₁-C₆)alkyl,(C₃-C₈)cycloalkyl, halo(C₁-C₆)alkyl, cyano, —C(O)R^(a), —CO₂R^(a),—C(O)NR^(a)R^(b), —SR^(a), —S(O)R^(a), —SO₂R^(a), —SO₂NR^(a)R^(b),nitro, —NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b), —NR^(a)SO₂NR^(a)R^(b), —OR^(a),—OC(O)R^(a), —OC(O)NR^(a)R^(b), heterocycloalkyl, aryl, heteroaryl,aryl(C₁-C₄)alkyl, or heteroaryl(C₁-C₄)alkyl; each R^(c) is independently—S(O)R^(a), —SO₂R^(a), —NR^(a)R^(b) —NR^(a)C(O)OR^(a), —NR^(a)SO₂R^(b),or —CO₂R^(a); and R^(a) and R^(b) are each independently hydrogen,(C₁-C₄)alkyl, (C₁-C₄)alkoxy(C₁-C₄)alkyl-, (C₃-C₁₀)cycloalkyl,heterocycloalkyl, aryl, aryl(C₁-C₄)alkyl-, heteroaryl(C₁-C₄)alkyl-, orheteroaryl, wherein any said cycloalkyl, heterocycloalkyl, aryl, orheteroaryl group is optionally substituted 1, 2, or 3 times,independently, by halogen, hydroxyl, (C₁-C₄)alkoxy, amino,—NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂, (C₁-C₄)alkyl, halo(C₁-C₄)alkyl,—CO₂H, —CO₂(C₁-C₄)alkyl, —CONH₂, —CONH(C₁-C₄)alkyl, —CON((C₁-C₄)alkyl)₂,—SO₂(C₁-C₄)alkyl, —SO₂NH₂, —SO₂NH(C₁-C₄)alkyl, or —SO₂N((C₁-C₄)alkyl)₂;or R^(a) and R^(b) taken together with the nitrogen to which they areattached represent a 5-8 membered saturated or unsaturated ring,optionally containing an additional heteroatom selected from oxygen,nitrogen, and sulfur, wherein said ring is optionally substituted 1, 2,or 3 times, independently, by (C₁-C₄)alkyl, halo(C₁-C₄)alkyl, amino,—NH(C₁-C₄)alkyl, —N((C₁-C₄)alkyl)₂, hydroxyl, oxo, (C₁-C₄)alkoxy, or(C₁-C₄)alkoxy(C₁-C₄)alkyl-, wherein said ring is optionally fused to a(C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring; or R^(a)and R^(b) taken together with the nitrogen to which they are attachedrepresent a 6- to 10-membered bridged bicyclic ring system optionallyfused to a (C₃-C₈)cycloalkyl, heterocycloalkyl, aryl, or heteroarylring; or a pharmaceutically acceptable salt thereof. 7-9. (canceled) 10.The compound according to claim 1, wherein R¹ and R² are eachindependently (C₁-C₄)alkyl, or a pharmaceutically acceptable saltthereof.
 11. The compound according to claim 6, wherein: R¹ and R² areeach methyl, independently (C₁-C₄)alkyl; R³ is hydrogen; R⁴ is selectedfrom the group consisting of (C₁-C₃)alkyl and halogen; R⁵ is—NR^(a)R^(b); R⁶ is hydrogen, halogen, (C₁-C₄)alkyl, or (C₁-C₄)alkoxy;R^(a) is azetidinyl, oxetanyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, or tetrahydropyranyl, each of which isoptionally substituted 1 or 2 times, independently, by (C₁-C₄)alkyl; orR^(a) is cyclopentyl or cyclohexyl, each of which is optionallysubstituted by amino, —NH(C₁-C₄)alkyl, or —N((C₁-C₄)alkyl)₂, and R^(b)is hydrogen or (C₁-C₄)alkyl; or a pharmaceutically acceptable saltthereof.
 12. The compound according to claim 1, wherein R³ is hydrogen,or a pharmaceutically acceptable salt thereof.
 13. (canceled)
 14. Thecompound according to claim 1, wherein R⁴ is selected from the groupconsisting of (C₁-C₃)alkyl and halogen, or a pharmaceutically acceptablesalt thereof. 15-16. (canceled)
 17. The compound according to claim 1,wherein R⁵ is selected from the group consisting of (C₃-C₆)alkoxy,(C₃-C₆)cycloalkyloxy-, heterocycloalkyloxy-, heterocycloalkyl,—NH((C₃-C₆)cycloalkyl), —N((C₁-C₃)alkyl)((C₃-C₆)cycloalkyl),—NH(heterocycloalkyl), and —N((C₁-C₃)alkyl)(heterocycloalkyl), whereinany said (C₃-C₆)alkoxy, (C₃-C₆)cycloalkyloxy-, heterocycloalkyloxy-,heterocycloalkyl, or (C₃-C₆)cycloalkyl is optionally substituted 1 or 2times, independently, by halogen, hydroxyl, (C₁-C₃)alkoxy, amino,—NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂, (C₁-C₃)alkyl,(C₁-C₃)alkoxy(C₁-C₃)alkyl-, amino(C₁-C₃)alkyl-,((C₁-C₃)alkyl)NH(C₁-C₃)alkyl-, ((C₁-C₃)alkyl)₂N(C₁-C₃)alkyl-,(C₃-C₈)cycloalkyl, cyano, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b),phenyl, or heteroaryl, or a pharmaceutically acceptable salt thereof.18. The compound according to claim 17, wherein R⁵ is selected from thegroup consisting of (C₃-C₆)alkoxy, (C₃-C₈)cycloalkyloxy-, andheterocycloalkyloxy-, each of which is optionally substituted byhydroxyl, (C₁-C₃)alkoxy, amino, —NH(C₁-C₃)alkyl, —N((C₁-C₃)alkyl)₂,(C₁-C₃)alkyl, —CO₂R^(a), —C(O)NR^(a)R^(b), —SO₂NR^(a)R^(b), phenyl, orheteroaryl, or a pharmaceutically acceptable salt thereof.
 19. Thecompound according to claim 17, wherein R⁵ is selected from the groupconsisting of cyclopentyloxy, cyclohexyloxy, pyrrolidinyloxy,piperidinyloxy, and tetrahydropyranyloxy, each of which is optionallysubstituted by hydroxyl, (C₁-C₃)alkoxy, amino, —NH(C₁-C₃)alkyl,—N((C₁-C₃)alkyl)₂, (C₁-C₃)alkyl, —CO₂R^(a), —C(O)NR^(a)R^(b),—SO₂NR^(a)R^(b), phenyl, furanyl, thienyl, pyrrolyl, imidazolyl,pyrazolyl, triazolyl, tetrazolyl, oxazolyl, thiazolyl, isoxazolyl,isothiazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl,pyrazinyl, or pyrimidinyl, wherein R^(a) is (C₁-C₄)alkyl orphenyl(C₁-C₂)alkyl and R^(b) is hydrogen or (C₁-C4)alkyl, or apharmaceutically acceptable salt thereof.
 20. The compound according toclaim 1, wherein R⁵ is —NR^(a)R^(b), or a pharmaceutically acceptablesalt thereof.
 21. The compound according to claim 20, wherein R^(a) isazetidinyl, oxetanyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, or tetrahydropyranyl, each of which isoptionally substituted 1 or 2 times, independently, by (C₁-C₄)alkyl, andR^(b) is hydrogen or (C₁-C₄)alkyl, or a pharmaceutically acceptable saltthereof.
 22. The compound according to claim 20, wherein R^(a) iscyclopentyl or cyclohexyl, each of which is optionally substituted byamino, —NH(C₁-C₄)alkyl, or —N((C₁-C₄)alkyl)₂, and R^(b) is hydrogen or(C₁-C₄)alkyl, or a pharmaceutically acceptable salt thereof.
 23. Thecompound according to claim 1, wherein R⁶ is selected from the groupconsisting of hydrogen, —SO₂(C₁-C₄)alkyl, halogen, (C₁-C₆)alkyl,(C₁-C₄)alkoxy, phenyl, heteroaryl, and cyano, wherein said phenyl orheteroaryl group is optionally substituted 1 or 2 times, independently,by (C₁-C₄)alkoxy, —NR^(a)R^(b), R^(a)R^(b)N(C₁-C₄)alkyl-,(C₁-C₄)alkylheterocycloalkyl-, halogen, (C₁-C₄)alkyl, (C₃-C₈)cycloalkyl,or heterocycloalkyl, or a pharmaceutically acceptable salt thereof. 24.The compound according to claim 23, wherein R⁶ is hydrogen, halogen,(C₁-C4)alkyl, or (C₁-C₄)alkoxy, or a pharmaceutically acceptable saltthereof.
 25. The compound according to claim 23, wherein R⁶ is hydrogen,fluorine, chlorine, or bromine, or a pharmaceutically acceptable saltthereof.
 26. The compound according to claim 1, wherein R⁷ is selectedfrom the group consisting of hydrogen, (C₁-C₆)alkyl, (C₃-C₆)cycloalkyl,phenyl, and heteroaryl, wherein said phenyl or heteroaryl group isoptionally substituted 1 or 2 times, independently, by (C₁-C₄)alkoxy,—NR^(a)R^(b), R^(a)R^(b)N(C₁-C₄)alkyl-, (C₁-C₄)alkylheterocycloalkyl-,halogen, (C₁-C₄)alkyl, (C3-C8)cycloalkyl, or heterocycloalkyl, or apharmaceutically acceptable salt thereof.
 27. The compound according toclaim 26, wherein R⁷ is hydrogen or (C₁-C₄)alkyl, or a pharmaceuticallyacceptable salt thereof.
 28. The compound according to claim 1, whereinR⁸ is selected from the group consisting of (C₄-C₆)alkyl,(C₄-C₆)cycloalkyl, heterocycloalkyl, and phenyl, wherein said(C₄-C₆)alkyl, (C₄-C₆)cycloalkyl, heterocycloalkyl, or phenyl isoptionally substituted 1 or 2 times, independently, by —OR^(a),—NR^(a)R^(b), —NHCO₂R^(a), (C₁-C3)alkyl, R^(a)R^(b)N(C₁-C₃)alkyl-,R^(a)O(C₁-C₃)alkyl-, —CO₂R^(a), —C(O)NR^(a)R^(b), or —SO₂NR^(a)R^(b), ora pharmaceutically acceptable salt thereof.
 29. The compound accordingto claim 1 which is:N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-4-methylthiophene-3-carboxamide;N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide;N4(5-amino-2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide;or2-bromo—N-((2,6-dimethyl-4-oxo-1,4-dihydropyridin-3-yl)methyl)-5-(((trans)-4-(dimethylamino)cyclohexyl)(ethyl)amino)-4-methylthiophene-3-carboxamide;or a pharmaceutically acceptable salt thereof.
 30. A pharmaceuticalcomposition comprising the compound or pharmaceutically acceptable saltthereof according to claim 1 and a pharmaceutically acceptableexcipient. 31-33. (canceled)